CN106231900A - Compound and using method thereof - Google Patents

Compound and using method thereof Download PDF

Info

Publication number
CN106231900A
CN106231900A CN201580020934.4A CN201580020934A CN106231900A CN 106231900 A CN106231900 A CN 106231900A CN 201580020934 A CN201580020934 A CN 201580020934A CN 106231900 A CN106231900 A CN 106231900A
Authority
CN
China
Prior art keywords
compound
alkyl
independently
formula
expression
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
CN201580020934.4A
Other languages
Chinese (zh)
Other versions
CN106231900B (en
Inventor
G·钱凯塔
R·M·勒米厄
S·曹
Y·丁
Z·叶
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Laboratoires Servier SAS
Original Assignee
Agios Pharmaceuticals Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Agios Pharmaceuticals Inc filed Critical Agios Pharmaceuticals Inc
Publication of CN106231900A publication Critical patent/CN106231900A/en
Application granted granted Critical
Publication of CN106231900B publication Critical patent/CN106231900B/en
Expired - Fee Related legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/08—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a carbon chain containing alicyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/50—Pyridazines; Hydrogenated pyridazines
    • A61K31/501—Pyridazines; Hydrogenated pyridazines not condensed and containing further heterocyclic rings
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D237/00—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings
    • C07D237/02—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings not condensed with other rings
    • C07D237/06—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members
    • C07D237/10—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D237/14—Oxygen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D237/00—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings
    • C07D237/02—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings not condensed with other rings
    • C07D237/06—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members
    • C07D237/10—Heterocyclic compounds containing 1,2-diazine or hydrogenated 1,2-diazine rings not condensed with other rings having three double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, e.g. ester or nitrile radicals, directly attached to ring carbon atoms
    • C07D237/20—Nitrogen atoms
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
    • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
    • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/08—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a carbon chain containing alicyclic rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
    • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
    • C07D403/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/02—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings
    • C07D405/12—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
    • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings
    • C07D409/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D409/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms
    • C07D409/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having sulfur atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings
    • C07D413/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
    • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/02—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings
    • C07D417/12—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D417/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00
    • C07D417/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and sulfur atoms as the only ring hetero atoms, not provided for by group C07D415/00 containing three or more hetero rings

Abstract

The compound that this document describes suppression transglutaminase and the compositions comprising described compound.The method that there is also described herein the compound using suppression transglutaminase in treatment of cancer.

Description

Compound and using method thereof
Claim of priority
This application claims the preferential of international application sequence No.PCT/CN2014/073812 submitted on March 21st, 2014 Power, it is incorporated by herein by quoting.
Background of invention
Cancerous cell depends on glycolysis and produces for the biosynthetic cellular energy of lipid and nucleotide and biology Chemical intermediate, and the majority of " normal " cell in adult tissue utilizes aerobic respiration.Between cancerous cell and normal cell This basic difference in terms of cellular metabolism is referred to as a watt berg's effect (Warburg Effect).Knot as this difference Really, the pyruvate produced via glycolytic pathway is converted to lactic acid rather than for producing acetyl-CoA, and finally produces The citrate utilized in normal tricarboxylic acid cycle.In order to compensate these energy variation and maintain tricarboxylic acid cycle, cancer is thin Born of the same parents depend on glutamine metabolism, and this metabolism realizes by raising glutaminase active.Utilization to this phenomenon Can realize by suppressing the glutaminase active of this rising.
Summary of the invention
This document describes the compound containing heterocycle, and pharmaceutically acceptable salt, solvate and hydrate.These are changed Compound can be used for such as treating disease described herein by the transglutaminase in suppression patient.Additionally provide and be included in The compositions (such as, pharmaceutical composition) of this compound provided together and these compositionss treatment such as with glutamine Enzyme abnormal function or glutaminase active raise the purposes in the method for relevant disease and illness (including such as cancer).
In one embodiment, it is provided that the compound of formula (I) or its pharmaceutically acceptable salt:
Wherein
X is the C being optionally substituted3-C7Cycloalkylidene；
Each W, Y and Z independently be-S-,-CH=,-CH=CH-,-CH=CR1-、-CR1=CR1-,-O-,-N=, Or-NH-, condition be at least one of W, Y and Z for-CH=,
Each W', Y' and Z' independently be-S-,-CH=,-CH=CH-,-CH=CR2-、-CR2=CR2-、-O-、-N =or-NH-, condition is that at least one of W', Y' and Z' is for-CH=；Condition be one of W, Y and Z be-CH=CH-,-CH= CR1-or-CR1=CR1-, and
When W is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and Z independently be-CH=or-N =；
When Y is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of W and Z independently be-CH=or-N =；
When Z is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and W independently be-CH=or-N =；
Condition be two of W', Y' and Z' for CH=CH-,-CH=CR2-or-CR2=CR2-, and
When W' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, the most each Y' and Z' independently be-CH=or-N =；
When Y' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of W' and Z' independently be-CH= Or-N=；
When Z' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of Y' and W' independently be-CH= Or-N=；
Each R1And R2Independently be-NH2、-N(R3)-C(O)-R4、-C(O)-N(R3)-R4、-N(R3)-C(O)-O-R4、-N (R3)-C(O)-N(R3)-R4Or-N (R3)-C(O)-SR4；
Each R3Independently be hydrogen, C1-6Alkyl or aryl；
Each R4Independently be C1-6Alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloalkyl, cycloalkyl-alkyl, Heterocyclylalkyl or heterocyclic radical, each of which R occurred by 0 to 3 time5Replace；
Each R5Independently be C1-6Alkyl, C1-6Alkoxyl ,-O-C1-6Alkylidene C1-6Alkoxyl ,-O-heterocyclic radical, C1-6Sulfur For alkoxyl, C1-6Haloalkyl, C3-7Cycloalkyl, C3-7Cycloalkyl-alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, heterocycle Alkyl, heterocyclic radical, cyano group, halogen, oxo ,-OH ,-OCF3、-OCHF2、-SO2-C1-6Alkyl ,-NO2、-N(R7)-C(O)-C1-6Alkane Base ,-C (O) N (R7)2、-N(R7)S(O)1-2-C1-6Alkyl ,-S (O)2N(R7)2、-N(R7)2、-C1-6Alkylidene-N (R7)2, wherein Described alkyl, C1-6Alkoxyl ,-O-C1-6Alkylidene C1-6Alkoxyl ,-O-heterocyclic radical, C1-6Thio alkoxy, C1-6Haloalkyl, C3-7Cycloalkyl, C3-7Cycloalkyl-alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloheteroalkylalkyl, heterocyclic radical ,-SO2- C1-6Alkyl ,-NO2、-N(R7)-C(O)-C1-6Alkyl ,-C (O) N (R7)2、-N(R7)S(O)1-2-C1-6Alkyl ,-S (O)2N(R7)2、- N(R7)2, or-C1-6Alkylidene-N (R7)2The R optionally occurred by 0 to 3 time8Replace；Or two R closed on5Part, with it The atom that connects collectively form cycloalkyl or heterocyclic radical；
Each R6Independently be hydrogen, fluorine, C1-6Alkyl ,-OH ,-NH2、-NH(CH3)、-N(CH3)2, or C1-6Alkoxyl；
Each R7Independently be hydrogen or C1-6Alkyl；
Each R8Independently be halogen, C1-6Alkyl, C1-6Haloalkyl ,-OH ,-N (R7)2, or C1-6Alkoxyl ,-O-C1-6 Alkylidene C1-6Alkoxyl, CN, NO2、-N(R7)-C(O)-C1-6Alkyl ,-C (O) N (R7)2、-N(R7)S(O)1-2C1-6Alkyl or-S (O)2N(R7)2；
M is 0,1 or 2；
N is 0,1 or 2；
O is 1,2 or 3；And
P is 1,2 or 3.
In another embodiment, it is provided that comprise the compound of formula (I) or the compositions of its pharmaceutically acceptable salt. In some embodiments, said composition is pharmaceutical composition.
In another embodiment, there is provided herein treatment or prevent as described herein (such as, treatment) disease, Illness or the method for disease, including using compound described herein, its pharmaceutically acceptable salt or comprising described herein Compound or the pharmaceutical composition of its pharmaceutically acceptable salt.
In another embodiment, there is provided herein the method such as suppressing transglutaminase in patient in need. In some embodiments, there is provided herein the product water of reduction transglutaminase in experimenter (patient the most in need) Flat.The method includes using the compound described herein of effective dose to experimenter in need or it is pharmaceutically acceptable Salt, thus suppresses the level of this experimenter's GLN enzyme.
In another embodiment, there is provided herein treatment suffer from or be susceptible to suffer from and experimenter's GLN in need Disease that the activity of the abnormal function of enzyme or the rising of transglutaminase is relevant or the method for the experimenter of disease.The method includes Use the compound described herein step to this experimenter in need of effective dose, thus treat, prevent or alleviate this and be subject to The disease of examination person or disease.In certain embodiments, said composition provides in pharmaceutical composition.In some embodiment In, the method includes identifying or select to will benefit from suppressing transglutaminase or reducing the experimenter of glutamine enzyme level.Example As, this experimenter can the level of cell or tissue sample GLN enzymatic activity based on this experimenter identify, it is used for controlling Treat the cancer relevant to abnormal transglutaminase function or activity.In another embodiment, selected experimenter for suffering from or Person is susceptible to suffer from disease or disease (disease being such as characterized, the such as cancer identified herein with the growth of undesirable cell or propagation Or other neoplastic condition) patient.
In another embodiment, there is provided herein the method for cancer in treatment experimenter, the method includes: optionally, Obtain Samples subjects；Obtain the assessment of this Samples subjects or assess this Samples subjects, wherein this Samples subjects It is characterised by that i) low-level CAM 120/80 is expressed compared with reference standard, ii) high-caliber waveform compared with reference standard Protein expression, or iii) low-level or drop low-level carboxylase expression of enzymes；And use to this experimenter in need The compound described herein of effective dose in treatment.In some embodiments, this Samples subjects is characterised by i) and reference Standard is compared low-level CAM 120/80 and is expressed and ii) high-caliber Vimentin compared with reference standard.At some In embodiment, this Samples subjects is characterised by or is additionally characterized by compared with reference standard low-level or drops low-level Carboxylase expression of enzymes.
In another embodiment, there is provided herein the method for cancer in treatment experimenter, this Subject characteristics is i) Low-level CAM 120/80 is expressed compared with reference standard, ii) high-caliber Vimentin compared with reference standard, or Iii) low-level or drop low-level carboxylase expression of enzymes；Including effective dose on experimenter's administering therapeutic in need Compound described herein.In some embodiments, this Subject characteristics is i) low-level E-compared with reference standard Cadherin is expressed and ii) high-caliber Vimentin compared with reference standard.In some embodiments, this experimenter It is characterised by or is additionally characterized by compared with reference standard low-level or drop low-level carboxylase expression of enzymes.
Detailed description of the invention
In following description statement or accompanying drawing shown in the structure of component and the details of arrangement be not intended to restricted 's.Embodiment can be implemented and carried out in various manners.Wording used herein and term also be in order at description purpose and not It is considered as restrictive." include " herein, " comprising " or " having ", " containing ", " relating to " and the use of version thereof It is intended to project listed after which and equivalent thereof and additional project.
Compound
This document describes compound and the compositions of suppression transglutaminase.The compound of suppression transglutaminase can be used In the disease treating such as neoplastic condition (such as cancer).
In one embodiment, it is provided that the compound of formula (I) or its pharmaceutically acceptable salt, or contained (I) compound or the pharmaceutical composition of its pharmaceutically acceptable salt:
Wherein
X is the C being optionally substituted3-C7Cycloalkylidene；
Each W, Y and Z independently be-S-,-CH=,-CH=CH-,-CH=CR1-、-CR1=CR1-,-O-,-N=or- NH-, condition be at least one of W, Y and Z for-CH=,
Each W', Y' and Z' independently be-S-,-CH=,-CH=CH-,-CH=CR2-、-CR2=CR2-、-O-、-N =or-NH-, condition is that at least one of W', Y' and Z' is for-CH=；Condition be one of W, Y and Z be-CH=CH-,-CH= CR1-or-CR1=CR1-, and
When W is CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and Z independently be-CH=or-N =；
When Y is CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of W and Z independently be-CH=or-N =；
When Z is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and W independently be-CH=or-N =；
Condition be two of W', Y' and Z' for CH=CH-,-CH=CR2-or-CR2=CR2-, and
When W' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, the most each Y' and Z' independently be-CH=or-N =；
When Y' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of W' and Z' independently be-CH= Or-N=；
When Z' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of Y' and W' independently be-CH= Or-N=；
Each R1And R2Independently be-NH2、-N(R3)-C(O)-R4、-C(O)-N(R3)-R4、-N(R3)-C(O)-O-R4、-N (R3)-C(O)-N(R3)-R4Or-N (R3)-C(O)-SR4；
Each R3Independently be hydrogen, C1-6Alkyl or aryl；
Each R4Independently be C1-6Alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloalkyl, cycloalkyl-alkyl, Heterocyclylalkyl or heterocyclic radical, each of which R occurred by 0 to 3 time5Replace；
Each R5Independently be C1-6Alkyl, C1-6Alkoxyl ,-O-C1-6Alkylidene C1-6Alkoxyl ,-O-heterocyclic radical, C1-6Sulfur For alkoxyl, C1-6Haloalkyl, C3-7Cycloalkyl, C3-7Cycloalkyl-alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, heterocycle Alkyl, heterocyclic radical, cyano group, halogen, oxo ,-OH ,-OCF3、-OCHF2、-SO2-C1-6Alkyl ,-NO2、-N(R7)-C(O)-C1-6Alkane Base ,-C (O) N (R7)2、-N(R7)S(O)1-2-C1-6Alkyl ,-S (O)2N(R7)2、-N(R7)2、-C1-6Alkylidene-N (R7)2, wherein Described alkyl, C1-6Alkoxyl ,-O-C1-6Alkylidene C1-6Alkoxyl ,-O-heterocyclic radical, C1-6Thio alkoxy, C1-6Haloalkyl, C3-7Cycloalkyl, C3-7Cycloalkyl-alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloheteroalkylalkyl, heterocyclic radical ,-SO2- C1-6Alkyl ,-NO2、-N(R7)-C(O)-C1-6Alkyl ,-C (O) N (R7)2、-N(R7)S(O)1-2-C1-6Alkyl ,-S (O)2N(R7)2、- N(R7)2, or-C1-6Alkylidene-N (R7)2The R optionally occurred by 0 to 3 time8Replace；Or two R closed on5Part, with it The atom that connects collectively form cycloalkyl or heterocyclic radical；
Each R6Independently be hydrogen, fluorine, C1-6Alkyl ,-OH ,-NH2、-NH(CH3)、-N(CH3)2, or C1-6Alkoxyl；
Each R7Independently be hydrogen or C1-6Alkyl；
Each R8Independently be halogen, C1-6Alkyl, C1-6Haloalkyl ,-OH ,-N (R7)2, or C1-6Alkoxyl ,-O-C1-6 Alkylidene C1-6Alkoxyl, CN, NO2、-N(R7)-C(O)-C1-6Alkyl ,-C (O) N (R7)2、-N(R7)S(O)1-2C1-6Alkyl or-S (O)2N(R7)2；
M is 0,1 or 2；
N is 0,1 or 2；
O is 1,2 or 3；And
P is 1,2 or 3.
In one embodiment, it is provided that the compound of formula (I) or its pharmaceutically acceptable salt, or contained (I) compound or the pharmaceutical composition of its pharmaceutically acceptable salt:
Wherein
X is the C being optionally substituted3-C7Cycloalkylidene；
Each W, Y and Z independently be-S-,-CH=,-CH=CH-,-CH=CR1-、-CR1=CR1-,-O-,-N=, Or-NH-, condition be at least one of W, Y and Z for-CH=,
Each W', Y' and Z' independently be-S-,-CH=,-CH=CH-,-CH=CR2-、-CR2=CR2-、-O-、-N =or-NH-, condition is that at least one of W', Y' and Z' is for-CH=；Condition be one of W, Y and Z be-CH=CH-,-CH= CR1-or-CR1=CR1-, and
When W is CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and Z independently be-CH=or-N =；
When Y is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of W and Z independently be-CH=or-N =；
When Z is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and W independently be-CH=or-N =；
Condition be two of W', Y' and Z' for CH=CH-,-CH=CR2-or-CR2=CR2-, and
When W' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, the most each Y' and Z' independently be-CH=or-N =；
When Y' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of W' and Z' independently be-CH= Or-N=；
When Z' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of Y' and W' independently be-CH= Or-N=；Each R1And R2Independently be-NH2、-N(R3)-C(O)-R4、-C(O)-N(R3)-R4、-N(R3)-C(O)-O-R4、-N (R3)-C(O)-N(R3)-R4Or-N (R3)-C(O)-SR4；
Each R3Independently be hydrogen, C1-6Alkyl or aryl；
Each R4Independently be C1-6Alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloalkyl, cycloalkyl-alkyl, Heterocyclylalkyl or heterocyclic radical, each of which R occurred by 0 to 3 time5Replace；
Each R5Independently be C1-6Alkyl, C1-6Alkoxyl, C1-6Thio alkoxy, C1-6Haloalkyl, C3-7Cycloalkyl, C3-7Cycloalkyl-alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloheteroalkylalkyl, heterocyclic radical, cyano group, halogen, oxo ,- OH、-OCF3、-OCHF2、-SO2-C1-6Alkyl ,-NO2、-N(R7)-C(O)-C1-6Alkyl ,-N (R7)2, or two R closed on5Portion Point, atom in connection collectively forms heterocyclic radical；
Each R6Independently be hydrogen, fluorine, C1-6Alkyl ,-OH ,-NH2、-NH(CH3)、-N(CH3)2, or C1-6Alkoxyl；
Each R7Independently be hydrogen or C1-6Alkyl；
M is 0,1 or 2；
N is 0,1 or 2；
O is 1,2 or 3；And
P is 1,2 or 3.
In some embodiments, X is unsubstituted cyclopropyl.In some embodiments, X is unsubstituted ring fourth Base.In some embodiments, X is unsubstituted cyclopenta.In some embodiments, X is cyclohexyl.Some embodiment party In case, X is suberyl.In some embodiments, X is replaced by 1 to 3 substituent group.In some embodiments, X is taken by 1 Replace for base.In some embodiments, X is replaced by 2 substituent groups.
In some embodiments, Y' is-N=.In some embodiments, Z' is-N=.In some embodiments, W' is-S-.In some embodiments, W' is-CH=CH-.In some embodiments, W is-CH=CH-.Implement at some In scheme, Y is-N=.In some embodiments, Z is-N=.In some aspects of these embodiments, W' is-S-, Y' It is-N=for-N=and Z'.In some aspects of these embodiments, W is-CH=CH-, and Y is-N=, and Z is-N=. In some aspects of these embodiments, W' is-CH=CH-, and Y' is-N=, and Z' is-N=.In these embodiments Some aspects, W is-CH=CH-, and Y is-N=, and Z is-N=, and W' is-S-, Y' be-N=and Z' be-N=.Implement at these Some aspects of scheme, W is-CH=CH-, and Y is-N=, and Z is-N=, and W' is-CH=CH-, Y' be-N=and Z' be-N=. In some embodiments, o is 1.In some embodiments, p is 1.In some embodiments, o is 1 and p to be 1.
In some embodiments, m is 0.In some embodiments, n is 0.In some embodiments, m is 0 and n It is 0.In some embodiments, R1And R2Identical.In some embodiments, R1And R2Different.
In some embodiments, m is 1.In some embodiments, n is 1.In some embodiments, n is 1 and m It is 1.In some aspects of these embodiments, each R6It is all hydrogen.In some embodiments, R1And R2Identical.Real at some Execute in scheme, R1And R2Different.
In some embodiments, R1And R2It is respectively-N (R3)-C(O)-R4, the most each R3For hydrogen and each R4For virtue Alkyl or heteroarylalkyl, each of which R occurred by 0 to 3 time5Replace.In some aspects of these embodiments, R1And R2Phase With.
In some embodiments, R1And R2It is respectively-N (R3)-C(O)-R4, the most each R3For hydrogen.These embodiment party Some aspects of case, each R4For the R occurred by 0 to 3 time5Substituted aralkyl.In some aspects of these embodiments, R1 And R2Identical.
In some aspects of these embodiments, each R4For the R occurred by 0 time5Substituted aralkyl (such as benzyl). In some aspects of these embodiments, each R4For the R occurred by 1 time5Substituted aralkyl (such as benzyl).Real at these Execute some other sides of scheme, each R5It is all-N (CH3)2.At the other aspect of these embodiments, each R5All For C1-6Alkoxyl (such as, methoxyl group or isopropoxy).At the other aspect of these embodiments, each R5It is all-O- Heterocyclic radical (such as ,-O-oxetanes).At the other aspect of these embodiments, each R5Be all halogen (such as, Fluorine or chlorine).At the other aspect of these embodiments, each R5It is all-NH2.Other side in these embodiments Face, each R5It is all-SO2-CH3.At the other aspect of these embodiments, each R5It is all-NHC (O) CH3.At these The other aspect of embodiment, each R5It is all-NO2.At the other aspect of these embodiments, each R5It is all Cyano group.At the other aspect of these embodiments, each R5It is all C1-6Halogenated alkoxy (such as, trifluoromethoxy).? The other aspect of these embodiments, each R5It is all C1-6Haloalkyl (such as, trifluoromethyl).These embodiment party The other aspect of case, each R5It is all C1-6Alkyl (such as, methyl).In some aspects of these embodiments, each R4 It is all by the R of twice appearance5Substituted aralkyl (such as benzyl).At some other sides of these embodiments, two R5For Halogen (such as, fluorine), another two R5For C1-6Alkoxyl (such as, methoxyl group).At the other aspect of these embodiments, Each R5It is all halogen (such as, fluorine).At the other aspect of these embodiments, each R5It is all C1-6Alkoxyl is (such as, Methoxyl group).At the other aspect of these embodiments, two R closed on5Part atom in connection is formed together Heterocyclic ring, obtains the part of following structure:
In some aspects of these embodiments, each R4It is all the R occurred by 0 to 3 time5Substituted heteroarylalkyl (example As, 2-pyridylmethyl, 2-pyridyl-ethyl group, 3-pyridylmethyl, 4-pyridylmethyl, 2-pyrazinyl-methyl, 2-thienyl Methyl, 2-indolylinethyl, 4-indolylinethyl, 2-Pyrimidylmethyl or 2-benzothiazolylmethyl).In these embodiments one A little aspects, each R4It is all the R occurred by 0 time5Substituted heteroarylalkyl (such as, 2-pyridylmethyl, 2-pyridyl-ethyl group, 3- Pyridylmethyl, 4-pyridylmethyl, 2-pyrazinyl-methyl, 2-thienyl methyl, 2-indolylinethyl, 3-indolylinethyl, 4-indolylinethyl, 2-Pyrimidylmethyl or 2-benzothiazolylmethyl).At the other side of these embodiments, each R4It is all The R occurred by 1 time5Substituted heteroarylalkyl (such as, 5-isoxazolyl, 2-pyridylmethyl or 3-indolylinethyl).At this Some other sides of a little embodiments, each R5It is all C1-6Alkyl (such as, methyl).These embodiments other its Its aspect, each R5It is all C1-6Alkoxyl (such as, methoxyl group).At the other aspect of these embodiments, each R5All For cyano group.At the other aspect of these embodiments, each R5It is all-N (CH3)2.These embodiments other its Its aspect, each R5It is all-NHC (O) CH3.At the other aspect of these embodiments, each R5Be all halogen (such as, Bromo).
In some aspects of these embodiments, each R4For the R occurred by 0 to 3 time5Substituted C1-6Alkyl (such as first Base, ethyl, n-pro-pyl or isopropyl).In some aspects of these embodiments, each R4It is all the R occurred by 0 time5Substituted C1-6Alkyl (such as methyl, ethyl, n-pro-pyl or isopropyl).At the other side of these embodiments, each R4It is all by 1 The R of secondary appearance5Substituted C1-6Alkyl (such as methyl, ethyl or the tert-butyl group).At some other aspects of these embodiments, Each R5It is all C1-6Thio alkoxy (such as, thiornethoxy group).At the other aspect of these embodiments, each R5All For C1-6Haloalkyl (such as, trifluoromethyl).At the other aspect of these embodiments, each R5It is all-OH.
In some aspects of these embodiments, each R4It is all the R occurred by 0 to 3 time5Substituted aryl (such as benzene Base).In some aspects of these embodiments, each R4It is all the R occurred by 0 time5Substituted aryl (such as benzyl).
In some aspects of these embodiments, each R4It is all the R occurred by 1 time5Substituted aryl (such as benzyl), Wherein R5For heterocyclic radical (such as, azetidinyl), and R5Replaced by the halogen (such as, fluorine) of twice appearance.
In some aspects of these embodiments, each R4It is all the R occurred by 0 to 3 time5Substituted heterocyclic radical is (such as, 3-tetrahydrofuran base).In some aspects of these embodiments, each R4It is all the R occurred by 0 time5Substituted heterocyclic radical base (such as, 3-tetrahydrofuran base).
In some aspects of these embodiments, each R4It is all the R occurred by 0 to 3 time5Substituted cycloheteroalkylalkyl (example As, 2-oxolane ylmethyl).In some aspects of these embodiments, each R4It is all the R occurred by 0 time5Substituted miscellaneous Cyclylalkyl (such as, 2-oxolane ylmethyl).In some aspects of these embodiments, each R4For the R occurred by 0 time5 Substituted cycloheteroalkylalkyl, and by following representation:
In some aspects of these embodiments, each R4It is all the R occurred by 0 to 3 time5Substituted cycloalkyl is (such as, Cyclopenta).In some aspects of these embodiments, each R4It is all the R occurred by 0 time5Substituted cycloalkyl (such as, ring penta Base).
In some aspects of these embodiments, each R4It is all the R occurred by 0 to 3 time5Substituted cycloalkyl-alkyl (example As, Cvclopropvlmethvl).In some aspects of these embodiments, each R4It is all the R occurred by 0 time5Substituted cycloalkyl alkane Base (such as, Cvclopropvlmethvl).
In some aspects of these embodiments, each R4It is all the R occurred by 0 to 3 time5Substituted C1-6Thiazolinyl is (such as, Vinyl).In some aspects of these embodiments, each R4It is all the R occurred by 1 time5Substituted C1-6Thiazolinyl (such as, second Thiazolinyl).At some other aspects of these embodiments, each R5It is all heteroaryl (such as, 2-pyridine radicals).
In some aspects of these embodiments, a R4For the R occurred by 0 time5Substituted C1-6Alkyl (such as, first Base), other R4For the R occurred by 1 time5Substituted heteroarylalkyl (such as, 3-indolylinethyl), wherein R5For C1-6Alkyl (such as, methyl).
In some aspects of these embodiments, a R4For the R occurred by 0 time5Substituted C1-6Alkyl (such as, first Base), other R4For the R occurred by 1 time5Substituted heteroarylalkyl (such as, 2-pyridine radicals), wherein R5For heterocyclic radical (such as, Azetidinyl), and R5The halogen substiuted (such as, fluorine) occurred by 2 times.
In some aspects of these embodiments, a R4For the R occurred by 0 time5Substituted C1-6Alkyl (such as, first Base), other R4For the R occurred by 1 time5Substituted heteroarylalkyl (such as, 3-indolylinethyl), wherein R5For C1-6Alkyl (such as, methyl).
In some aspects of these embodiments, a R4For C1-6Alkyl (such as, methyl), other R4For heteroarylalkyl (such as, 2-pyridylmethyl), the R that each of which is occurred by 0 time5Replace.
In some aspects of these embodiments, a R4For the R occurred by 0 time5Substituted heteroarylalkyl (such as, 2-pyrrole Piperidinyl base), other R4For the R occurred by 1 time5Substituted aralkyl (such as, benzyl), wherein R5For C1-6Alkoxyl is (such as, Methoxyl group).
In some aspects of these embodiments, a R4For the R occurred by 0 time5Substituted C1-6Alkyl (such as, first Base), other R4For the R occurred by 1 time5Substituted aralkyl (such as, benzyl), wherein R5For C1-6Alkoxyl (such as, methoxy Base).
In some embodiments, R2For-NH2, and R1For-N (R3)-C(O)-R4, wherein R3For hydrogen and R4For being gone out by 0 time Existing R5Substituted heteroarylalkyl (such as, 2-pyridylmethyl).
In some embodiments, each R6It is all H.
In some embodiments, the compounds represented of the formula (II) of formula (I):
Wherein R1、R2、R3、R4、R5、R6, o, p, m, n and X be as defined in formula (I).
In some embodiments, the compounds represented of the formula (III) of formula (I):
Wherein R1、R2、R3、R4、R5、R6, o, p, m, n and X be as defined in formula (I).
In some embodiments, the compounds represented of the formula (IIa) of formula (I) or (II):
Wherein R1、R2、R3、R4、R5、R6, o, p, m, n and X be as defined in formula (I).
In some embodiments, the compounds represented of the formula (IIIa) of formula (I) or (III):
Wherein R1、R2、R3、R4、R5、R6, o, p, m, n and X be as defined in formula (I).
In some embodiments, the compounds represented of the formula (IIb) of formula (I) or (II) or (IIa):
Wherein R1、R2、R3、R4、R5、R6, o, p, m, n and X be as defined in formula (I).
In some embodiments, the compounds represented of the formula (IIIb) of formula (I) or (III) or (IIIa):
Wherein R1、R2、R3、R4、R5、R6, o, p, m, n and X be as defined in formula (I).
In some embodiments, the compounds represented of the formula (IV) of formula (I):
Wherein R1、R2、R3、R4And R5As defined in formula (I), and q is 0,1,2,3 or 4.
In some embodiments, the compounds represented of the formula (V) of formula (I):
Wherein R1、R2、R3、R4And R5As defined in formula (I), and q is 0,1,2,3 or 4.
In some embodiments, the compounds represented of the formula (IVa) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (Va) of formula (I) or (V):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (IVb) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (Vb) of formula (I) or (V):
Wherein R4、R5With q as defined in formula (V).In some embodiments, formula (I) or the formula of (IV) (IVc) compounds represented:
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (Vc) of formula (I) or (V):
Wherein R4、R5With q as defined in formula (V).
In some embodiments, the compounds represented of the formula (VIa) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (VIIa) of formula (I) or (V):
Wherein R4、R5With q as defined in formula (V).In some embodiments, formula (I) or the formula of (IV) (VIb) compounds represented:
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (VIIb) of formula (I) or (V):
Wherein R4、R5With q as defined in formula (V).
In some embodiments, the compounds represented of the formula (VIc) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (VIIc) of formula (I) or (V):
Wherein R4、R5With q as defined in formula (V).
In some embodiments, the compounds represented of the formula (VIIIa) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (IXa) of formula (I) or (V):
Wherein R4、R5With q as defined in formula (V).
In some embodiments, the compounds represented of the formula (VIIIb) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (IXb) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (VIIIc) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In some embodiments, the compounds represented of the formula (IXc) of formula (I) or (IV):
Wherein R4、R5With q as defined in formula (IV).
In certain embodiments, the compound during the exemplary compounds of Formulas I includes table 1 and described in embodiment.Example Algoscopy as described in by embodiment, can test the ability of its suppression transglutaminase to compound described herein. In table 1, the exemplary compounds of display is as follows.
Table 1:
The most also include the compound of formula I or the method for the compound of any embodiment described herein, bag Include and make
WithReaction, wherein G is Leaving group, such as halogen ,-OH, perfluoro alkyl sulfonic acid root (such as, trifluoromethanesulfonic acid root), tosylate, methanesulfonate, Or NH2。
In some embodiments, preceding method includes making
React to produceStep (1)；And makeThe step (2) of reaction.
In some embodiments, the compound of the compound of formula I or any embodiment described herein Method includes making
WithReaction, Wherein G is leaving group, such as halogen ,-OH, perfluoro alkyl sulfonic acid root (such as, trifluoromethanesulfonic acid root), tosylate, first Sulfonate radical or NH2。
In some embodiments, preceding method includes makingWithReact with ProduceStep (1)；
MakeWithReact to produce
Step (2) and makeWithThe step (3) of reaction.
Compound described herein can use multiple synthetic technology to prepare, described in example the most provided herein Those technology.As technical staff is to understand, the method for the synthesis compound of multiple formulas herein is for ordinary skill Personnel will be apparent from.It addition, each synthesis step can be carried out by substituting order or sequence, to obtain desired chemical combination Thing.Can be used for synthesizing the synthesis chemical conversion of compound described herein and blocking group method (protection and deprotection) in ability Territory is known, and such as including described in the documents below: R.Larock, Comprehensive Organic Transformations, VCH publishing house (1989)；T.W.Greene and P.G.M.Wuts, Protective Groups in Organic Synthesis, second edition, John Wiley and Sons (1991)；L.Fieser and M.Fieser,Fieser and Fieser's Reagents for Organic Synthesis,John Wiley and Sons(1994)；And L.Paquette compiles, Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995) and later release thereof.
Compound provided herein can contain one or more asymmetric centers, and therefore with racemate and raceme Presented in mixture, single enantiomer, independent non-corresponding isomer and non-corresponding isomer mixture.These are changed In the range of these type of isomeric form all of compound are expressly included in.Unless otherwise directed, otherwise when compound be by knot Structure is named or describes and do not indicate spatial chemistry, and when there is one or more chiral centre, it is thus understood that represent this change The all possible stereoisomer of compound.Compound provided herein can also containing can limit key rotate (such as, by ring or The existence of double bond and the restriction that causes) key (such as, carbon-carbon bond).Therefore, all of cis/trans and E/Z isomer are the brightest Really it is included.
Compound provided herein (such as, the compound of Formulas I) can also comprise one or more isotope and replace.Citing For, H can in any isotope form, including1H、2H (D or deuterium) and3H (T or tritium)；C can be in any isotope shape Formula, including12C、13C and14C；O can in any isotope form, including16O and18O；Etc..Compound provided herein is also Can represent by multiple tautomeric form, in these cases, include clearly compounds described herein all mutually Becoming isomeric form, even if single tautomeric form may be illustrate only, (such as, the alkylation of loop systems can cause multiple position The alkylation of point；In all these product are expressly included in).All these isomeric form of these compounds are the brightest Really it is included.In all crystal forms of compound described herein are expressly included in.
Compound provided herein includes these compounds itself, and its salt and its prodrug (if appropriate).Lift For example, salt can be formed between the substituent group (such as, amino) of positively charged in anion and compound described herein.Suitable Close anion include chloride ion, bromide ion, iodide ion, sulfate radical, nitrate anion, phosphate radical, citrate, methanesulfonate, three Fluoroethanoic acid root and acetate.Similarly, can electronegative substituent group (example in cation and compound described herein Such as, carboxylate radical) between form salt.Be suitable for cation include sodium ion, potassium ion, magnesium ion, calcium ion and ammonium sun from Son, such as trimethyl ammonium ion.The example of prodrug includes ester and other pharmaceutically acceptable derivates, and they are being executed to experimenter Reactive compound can be provided after with.
Compound provided herein can be modified by additional suitable functional group, biological with selected by strengthening Matter, such as, targeting particular organization.These modifications are well known in the art, and enter given biological compartment (example including increasing As, blood, lymphsystem, central nervous system) biological penetration, increase oral administration biaavailability, increase dissolubility with permit Permitted used by injection, change metabolism and change discharge rate those.
In an alternative embodiment, compound described herein can serve as platform or skeleton, these platforms or skeleton Can be used in combinatorial chemistry technique to prepare derivant and/or the chemical libraries of compound.The derivant of these compounds and Storehouse has biological activity and can be used for differentiating and design the multiple compound with given activity.It is suitable for use with described herein The combination technique of compound be well known in the art, such as Obrecht, D. and Villalgrodo, J.M., Solid- Supported Combinatorial and Parallel Synthesis of Small-Molecular-Weight Compound Libraries, Pergamon-Elsevier Science Limited (1998) are illustrated, including such as " split and merge (split and pool) " or " parallel " synthetic technology, solid phase and solution phase techniques, and coding techniques (ginseng See, such as, Czarnik, A.W., Curr.Opin.Chem.Bio., (1997) 1,60) etc..Therefore, an embodiment relates to Compound described herein is used to produce derivant or the method for chemical libraries, including: 1) main body including multiple hole is provided；2) to Each hole is provided one or more compounds identified by method described herein；3) in each hole, provide other one Individual or multi-chemical；4) one or more obtained products are isolated from each hole.Alternate embodiment is directed to use with this The compound that literary composition describes produces derivant or the method for chemical libraries, including: 1) provide one or more to be connected to solid support Compound described herein；2) one or more are connected to solid support to treat this with one or more other chemicals The compound identified by method described herein；3) isolate obtained one or more from this solid support to produce Thing.In the method being described above, can by " label " mark or mark part be connected to compound described herein or its Derivant and/or separate from compound described herein or derivatives thereof, to promote desired product or its intermediate Follow the trail of, identify or separate.These parts are known in this field.Chemicals used in method mentioned above is permissible Including such as solvent, reagent, catalyst, blocking group and Deprotection reagent etc..The example of these chemicals is this paper institute Those occurred in the different synthesis mentioned and blocking group Chemistry Teaching Material and paper.
Definition
Term " halogen " or " halogen " refer to any group in fluorine, chlorine, bromine or iodine.
Term " alkyl " refers to that it can be straight or branched containing the saturated or aliphatic unsaturated hydrocarbon specifying carbon number. For example, C1-C12Alkyl indicates this group can have 1 to 12 (including end points) carbon atom wherein.Term " alkane Base " include alkenyl part.Term " haloalkyl " refer to one or more hydrogen atom by the alkyl of halogen displacement, including all hydrogen all By the moieties (such as perfluoroalkyl) of halogen displacement.Term " aryl alkyl " or " aralkyl " refer to that alkyl hydrogen atom is fragrant The moieties of base group displacement.Aralkyl includes the group replaced by aromatic yl group more than a hydrogen atom." aryl alkane Base " or the example of " aralkyl " include benzyl, 2-phenylethyl, 3-phenyl propyl, 9-fluorenyl, benzhydryl and trityl Group.
Term " alkylidene " or " ring alkylidene " refer to divalent alkyl or cycloalkyl, such as-CH2-、-CH2CH2-and- CH2CH2CH2-。
Term " thiazolinyl " refers to straight chain or the branched hydrocarbon chains containing 2 to 12 carbon atoms and having one or more double bond.Alkene The example of base group includes but not limited to, pi-allyl, acrylic, crotyl, 3-hexenyl and 3-octenyl.The one of double key carbon The individual junction point that can be optionally alkenyl group.
Term " alkoxyl " refers to-O-alkyl group.Term " halogenated alkoxy " refers to that one or more hydrogen atom is put by halogen The alkoxyl changed, including all hydrogen the most by the alkoxy portion (such as perfluoro alkoxy) of halogen displacement.
Term " aryl " refers to monocycle, dicyclo or tricyclic aromatics loop systems, and any annular atoms wherein can being replaced is all (such as, being substituted with one or more substituents) can be replaced.The example of aryl moiety includes but not limited to, phenyl, naphthyl and Anthryl.Except as otherwise noted, otherwise any annular atoms in aryl all can be substituted with one or more substituents.
As used herein, term " cycloalkyl " includes ring-type, dicyclo, three rings or multi-ring non-aromatic with 3 to 12 carbon Groups.Any commutable annular atoms all can be replaced (such as, being substituted with one or more substituents).Group of naphthene base can Containing fused rings or volution.Fused rings is the ring of total common carbon atom.The example of cycloalkyl moiety includes but not limited to, ring third Base, cyclohexyl, methylcyclohexyl, adamantyl and norcamphanyl.
As used herein, term " cycloalkyl-alkyl " refers to the alkyl being substituted by cycloalkyl.
Term " heterocyclic radical " or " heterocyclic group " refer to 3-to 14-unit non-aromatic ring structure (such as, 3-to 14-ring, more excellent Elect 3-to 7-ring as), its ring structure includes one to four hetero atom independently selected from O, N and S.Heterocyclic radical or heterocyclic radical Fused rings or volution can be contained in group.Heterocycle can also be multi-ring, and each group has such as 5-7 ring members.Term " heterocyclic radical " Or " heterocyclic group " includes saturated and fractional saturation heterocyclyl structures.Hetero atom is optionally the company of heterocyclyl substituent Contact.
Term " heteroaryl " refer to have 1 to 3 ring hetero atom (if monocycle), 1 to 6 ring hetero atom (if Dicyclo) or 1 to 9 ring hetero atom (if three rings) 5-14 unit (that is, 5-8 unit monocycle, 8-12 unit dicyclo or 11-14 unit three rings) aromatic rings system, described ring hetero atom independently selected from O, N and S (such as, if being monocycle, double respectively Ring or three rings, then have 1-3,1-6 or 1-9 ring hetero atom N, O or S).Any commutable annular atoms all can be replaced (such as, being substituted with one or more substituents).
Containing one or more hetero atoms and fragrance with non-aromatic ring (wherein from loop systems to molecule remainder Junction point be by non-aromatic ring) dicyclo and the loop systems of three rings be considered as heterocyclic radical.Aryl or heteroaryl and ring Alkyl or heterocyclic radical condense and from loop systems to the junction point of molecule remainder be the dicyclo by aromatic rings or three ring rings System is considered as aryl or heteroaryl groups.
As used herein, term " cycloheteroalkylalkyl " refers to the alkyl replaced by heterocyclic group.
As used herein, term " heteroarylalkyl (hetaralkyl) " and " heteroarylalkyl (heteroaralkyl) " refer to by The substituted alkyl of heteroaryl groups.The ring hetero atom of compound provided herein includes N-O, S (O) and S (O)2。
Except as otherwise noted, otherwise aryl, heteroaryl, cycloalkyl and heterocyclyl groups, either single or conduct A part (such as, the aryl moiety of aralkyl) for group, is the most optionally replaced at one or more commutable atoms Base replaces, and these substituent groups are independently selected from halogen ,-C ≡ N, C1-C4Alkyl ,=O ,-ORb、-ORb’、-SRb、-SRb’、-(C1- C4Alkyl)-N (Rb)(Rb)、-(C1-C4Alkyl)-N (Rb)(Rb’)、-N(Rb)(Rb)、-N(Rb)(Rb’)、-O-(C1-C4Alkyl)-N (Rb)(Rb)、-O-(C1-C4Alkyl)-N (Rb)(Rb’)、-(C1-C4Alkyl)-O-(C1-C4Alkyl)-N (Rb)(Rb)、-(C1-C4Alkane Base)-O-(C1-C4Alkyl)-N (Rb)(Rb’)、-C(O)-N(Rb)(Rb)、-(C1-C4Alkyl)-C (O)-N (Rb)(Rb)、-(C1-C4 Alkyl)-C (O)-N (Rb)(Rb’)、-ORb’、Rb’、-C(O)(C1-C4Alkyl) ,-C (O) Rb’、-C(O)N(Rb’)(Rb)、-N(Rb)C (O)(Rb)、-N(Rb)C(O)(Rb’)、-N(Rb)SO2(Rb)、-SO2N(Rb)(Rb)、-N(Rb)SO2(Rb’) and-SO2N(Rb) (Rb’), any of which alkyl substituent is the most optionally by-OH ,-O-(C1-C4Alkyl), halogen ,-NH2、-NH(C1-C4Alkyl) Or-N (C1-C4Alkyl)2In one or more be further substituted with；
Each RbIndependently selected from hydrogen and-C1-C4Alkyl；Or
Two RbForming 4-to 8-unit heterocyclic radical together with the nitrogen-atoms being connected with them, this heterocyclic radical optionally comprises one Other hetero atom selected from N, S and O；And
Each Rb’Independently selected from C3-C7Carbocylic radical, phenyl, heteroaryl and heterocyclic radical, wherein at described phenyl, cycloalkanes One or more on base, heteroaryl or heterocyclic substituent may replace position optionally by-(C1-C4Alkyl) ,-(C1-C4Halothane Base) ,-OH ,-O-(C1-C4Alkyl) ,-O-(C1-C4Fluoroalkyl), halogen ,-NH2、-NH(C1-C4Alkyl) or-N (C1-C4Alkyl)2 In one or more be further substituted with.
Heterocyclyl groups, the either single or part as group, the most optionally one or more any By epoxide (=O) ,-C on commutable nitrogen-atoms1-C4Alkyl or the substituted C of fluorine1-C4Alkyl is replaced.
Term " is replaced " and refers to that hydrogen atom is by another group displacement.
Term " optionally " pointer to the inhibitory action of transglutaminase than other target exceed at least 2 times, 3 times, 4 Again, 5 times, 6 times or 10 times.
As used herein, term " inhibitor " instigates the enzymatic activity of transglutaminase measurably to slow down, stop, reducing Or the reagent of the level that inactivation is to decrease below transglutaminase normal activity level.Glutamine enzyme inhibitor can be peptide Or nucleic acid (such as glutamic acid).Can be by measuring the transglutaminase activity when accepting this agent treated directly or indirectly Assess to determine if it is inhibitor.The activity of this reagent can be measured by such as relative comparison material.In some cases, The measured activity of reagent is the inhibitory action for transglutaminase.
As used herein, term " is obtained " and refers to be obtained by " directly obtaining " or " indirectly obtaining " physical entity or value Must be to the proprietary rights of this physical entity maybe this value (such as digital value)." directly obtain " process of implementing that refers to (such as, to implement to close Become or the method for analysis) obtain this physical entity or value." indirectly obtain " and refer to (the most directly obtain from the opposing party or source Third party's laboratory of this physical entity or value) receive this physical entity or value.Directly obtain physical entity to include implementing one Process, this process includes the physical change of physical material (such as, raw material).Exemplary variations includes by two or more raw materials Prepare physical entity；Shear or fragmentation material；Isolated or purified material；By mixed for the synthesis of two or more separate group of entities Compound；Carrying out chemical reaction, this chemical reaction includes fracture or forms covalently or non-covalently key.Directly acquired value includes implementing One process including sample or the additionally physical change of material, such as, implement to include material (such as sample, analyte or reagent) The analysis process (sometimes herein called " physical analysis ") of middle physical change, implements analysis method, such as, includes following one Kind or multiple method: from another kind of material isolated or purified one material, such as analyte or fragment or it other derive Thing；By analyte or its fragment or other derivant and another kind of material (such as buffer, solvent or reactant) combination； Or change analyte or its fragment or the structure of other derivant, such as by first and second atom in this analyte Between rupture or form covalently or non-covalently key；Or by changing reagent or its fragment or the structure of other derivant, such as By rupturing between first and second atom of this reagent or forming covalently or non-covalently key.
When " acquisition sample " term is as used herein, refer to that sample is (such as, by " directly obtaining " or " indirectly obtaining " Tissue sample or nucleic acid samples) obtain the proprietary rights of this sample." directly obtain sample " and be meant to implement a process (example As, implement physical method, such as operation or extraction) obtain this sample." indirectly obtain sample " and refer to from the opposing party or source (example As directly obtained third party's laboratory of this sample) receive this sample.Directly obtain sample to include implementing a process, this mistake Journey includes the physical change in physical material, this physical material for example, raw material, such as tissue, such as tissue in people patient or it The front tissue separated from patient.Exemplary variations includes being prepared physical entity by raw material, dissects or scrapes undertissue；Isolated or purified Material (such as sample tissue or nucleic acid samples)；By two or more separate combination of entities resulting mixtures；Carry out including breaking Split or formed the chemical reaction of covalently or non-covalently key.Directly obtaining sample to include implementing a process, this process includes sample Or the physical change in another material, such as, as described above.
As used herein, the CAM 120/80 expression of " relatively low " refers to thin at epithelium with CAM 120/80 compared with reference standard Expression in born of the same parents compares relatively low, reduction or non-existent CAM 120/80 expression, as by side as known in the art Any one in method, such as documents below is characterized: (Yauch etc., (2005) Clin Cancer Res 11:24； Savagner etc., (2010) Ann Oncol.21 (suppl7): vii89；Thiery etc., (2002) Nature Reviews Cancer 2 (6): 442).
As used herein, the Vimentin level of " higher " refers to Vimentin in epithelial cell compared with reference standard Expression compare Vimentin level that is higher or that increase, as by method as known in the art, such as with hereafter Any one in offering is characterized: (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (suppl7): vii89；Thiery etc., (2002) Nature Reviews Cancer 2 (6): 442).
As used herein, relatively low compared with reference standard " or the pyruvate carboxylase expression of " reduction " refer to as by The expression in epithelial cell of any one CAM 120/80 characterized in method as known in the art, such as documents below Level compares relatively low, reduction or non-existent CAM 120/80 expression: (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (suppl7): vii89；Thiery etc., (2002) Nature Reviews Cancer 2 (6): 442).
As used herein, " cancer " and " tumor " are the terms of synonym.Term " cancer " or " tumor " refer to have carcinogenic carefully The existence of the typical characteristic cell of born of the same parents, as uncontrolled propagation, immortality, metastatic potential, quickly growth and multiplication rate with And some characteristic morphologic feature.Cancerous cell often form in tumor, but these cells can separately exist in animal body, Can be maybe non-tumorigenic cancer cells, such as leukaemia.These cells can have the typical characteristic of mesenchymal cell, as following Any one in document is characterized: (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (supplementary issue 7): vii89；Thiery etc., (2002) Nature Reviews Cancer 2 (6): 442).
Abbreviation Me, Et, Ph, Tf, Nf, Ts, Ms represent methyl, ethyl, phenyl, trifluoromethane sulfonyl group, nine fluorine fourths respectively Alkane sulfonyl, p-toluenesulfonyl and methane sulfonyl.This area has the abbreviation that the organic chemist of common skill is utilized More fully inventory see Journal of Organic Chemistry (Journal of Organic Chemistry) each volume the first phase in； This inventory typically appear in entitled standardized abbreviations inventory (Standard List of Abbreviations) table in.Institute All abbreviations that stating has the organic chemist of common skill to be utilized in abbreviation contained in inventory and this area are all passed through to draw With being expressly incorporated herein.
The method of assessment compound
Glutaminase active can be monitored by the generation of detection product glutamic acid or ammonia.In some embodiments In, measure the generation of glutamic acid, because ammonia is any one product of multiple biological respinse.
Glutamic acid produce can be measured by any one in multiple standards method as known in the art, such as chemistry and Chromatographic detection method, and utilize the conjugate enzyme of NADH and glutamte dehydrogenase to measure.Extracellular glutamate concentration can also be Internal use microdialysis as known in the art method is measured.It is based on micro-for measuring a kind of applicable method of glutamic acid Two step measurements of amount titration, in this mensuration, glutamte dehydrogenase makes the glutamic acid deamination quantitatively formed in initial step Base, obtains the NADH (Godfrey) etc. of equivalent, and 1977；Kvamme etc., 1985), can detect with spectrophotography subsequently.
Therapeutic Method
In one embodiment, it is provided that for treating or preventing the disease of (such as treatment), disease as described herein Suffer from or the method for disease, including administered compound, the pharmaceutically acceptable salt of compound or comprise compound described herein The pharmaceutical composition of (such as, the compound in formula (I) or table 1).
Compound described herein and compositions can be used to the cell the most in vitro or in isolated culture, or such as exist Internal be applied to experimenter, with treatment, prevent and/or diagnose various disease conditions, including the most described below those.
As used herein, term " treatment (treat/treatment) " is defined as applying or using individually or with second The united compound of compound to experimenter, such as patient；Or apply or use this compound to from suffering from disease (such as, As described herein disease), the symptom with disease or the tissue having the experimenter (such as patient) of tendency of ill disease to separate Or cell, such as cell line, with reach to cure, cure, alleviate, alleviate, change, correct, improve, improve or affect this disease, One or more symptoms of this disease or the tendency suffering from this disease (such as, prevent at least one symptom of this disease or delay this The outbreak of at least one symptom of disease) purpose.
As used herein, the amount of effective sanatory compound, or " therapeutically effective amount " refer to single or multiple agent Amount at treatment cell, or is cured, alleviates, alleviates or is improved and suffer from the experimenter of disease and exceed and do not exist after experimenter uses The amount of the compound of situation desired in the case of this treatment.
As used herein, effectively prevent the amount of the compound of disease, or " effective dose in prevention " of compound refers to list Secondary or multidose effectively prevents or delays outbreak or the appearance of recurrence of the symptom of disease or this disease after experimenter uses Amount.
It is the as used herein of synonym with " experimenter " as used herein the term " patient ", refers to become or have become as Treatment, the animal of object observed and/or test, typically the mankind (that is, the sex of any age cohort, such as Section patient or adult patients), or other mammal, such as primate (such as machin, Rhesus Macacus)；Commercial relevant Mammal, such as cattle, pig, horse, sheep, goat, cat and/or Canis familiaris L.；And/or birds, including commercial relevant birds, as chicken, Duck, goose and/or turkey.When this term and compound or medicine use be used in combination time, then this patient has become as treatment, sees Examine and/or this compound or the object of medicament administration.
Cancer
Method described herein may be used for any cancer, such as national cancer institute (National Cancer Institute) those described by.Can be estimated cancer determining whether it uses method described herein.Exemplary Cancer may include but be not limited to pulmonary carcinoma, such as nonsmall-cell lung cancer；Breast carcinoma, such as three negative breast cancer；Or hepatocarcinoma, bone Sarcoma, lipoma, chondrosarcoma or mesothelioma.In some embodiments, this cancer is selected from colon cancer, renal cell carcinoma, acute Myelomatosis (AML), melanoma and multiple myeloma.
This cancer can be primary tumor, is i.e. positioned at the anatomical site that tumor growth is initial.This cancer can also be Transfer, i.e. occur at least the second anatomical site being different from the initial anatomical site of tumor growth.This cancer can be Relapse cancer, i.e. the cancer after the treatment and reappeared after this cancer detection is less than a period of time.Relapse cancer can solve Cut open and on, be positioned primary tumor partly, the most anatomically near primary tumor；Regionally it is positioned primary tumor, Such as it is positioned in the lymph node near primary tumor；Or it is located remotely from primary tumor, it is located remotely from former the most anatomically In the region of beginning tumor.
Cancer combination treatment
In some embodiments, compound described herein is to execute together with the treatment of cancer other with one or more With.Exemplary cancers treatment includes, such as: chemotherapy, targeted therapies (such as antibody therapy), immunotherapy and hormonotherapy. Under these each examples for the treatment of are provided in.
Chemotherapy
In some embodiments, compound described herein is to use together with one or more chemotherapy.Chemistry Therapy is with destroying the treatment of cancer that the medicine of cancerous cell is carried out." chemotherapy " impact the most generally quickly division The cytotoxic drug of cell, is contrasted with targeted therapies.Chemotherapy drugs divides with various possible mode interference cells Split, the separation of the chromosome replicating or being newly formed of such as interference DNA.Chemotherapeutic most of form all targeting are all quickly The cell of division, is not specific to cancerous cell, but specificity to a certain degree may be from can not repairing in many cancerous cell DNA damage, and normal cell is the most permissible.
The example of chemotherapeutant used in cancer therapy includes such as antimetabolic species (such as, folic acid, purine and phonetic Piperidine derivatives) and alkylating agent class (such as, nitrogen mustards, nitrosoureas, platinum, alkyl sulfonates, hydrazine, Triazenes, nitrogen Third pyridine class, spindle poison, cytotoxic agent class, topoisomerase enzyme inhibitor class and other).Exemplary Agents includes A Rou ratio Star, D actinomycin D, alitretinoin (Alitretinon), hexamethyl melamine, aminopterin, aminolevulinic acid, amrubicin, peace Acridine, anagrelide, arsenic trioxide, asparaginase, atrasentan, Belotecan, Bexarotene, bendamustine, rich Bleomycin, bortezomib, busulfan, camptothecine, capecitabine, carboplatin, carboquone, carmofur, carmustine, celecoxib, Chlorambucil, chlormethine, cisplatin, cladribine, clofarabine, Ke Lita enzyme, cyclophosphamide, cytosine arabinoside, dacarbazine, more Mildew element, daunomycin, decitabine, Demecolcine, Docetaxel, doxorubicin, Efaproxiral, department of Erie not, according to sand Lu Xing, enocitabine, epirubicin, estramustine, etoglucid, etoposide, floxuridine, fludarabine, fluorouracil (5FU), fotemustine, gemcitabine, Ge Li get implant, hydroxyurea (Hydroxycarbamide), hydroxyurea (Hydroxyurea), idarubicin, ifosfamide, irinotecan, Yi Luofufen, ipsapirone, La Luotasai, folinic acid, Mycocet, daunomycin liposome, lonidamine, lomustine, lucanthone, mannomustine, masoprocol, American and French Logical sequence, purinethol, mesna, methotrexate, MAL, mitobronitol, mitoguazone, mitotane, mitogen are mould Element, mitoxantrone, nedaplatin, nimustine, Ao Limosen, Ao Maxitaxin, Ortataxel, oxaliplatin, Pacific Ocean purple China fir alcohol, pegaspargase, pemetrexed, pentostatin, Pirarubicin, a China fir fine jade, plicamycin, porfimer sodium, prednimustine, Procarbazine, thunder are for Qu Sai, Reynolds Mo Siting, Rubitecan, 1-(2-C-cyano-2-dioxy-BETA-D-arabino-pentofuranosyl)-N4-palmitoyl cytosine, semustine, Sai Xima collection, Satraplatin JM216 BMS 182751, chain assistant Star, talaporfin, Tegafur-uracil mixt., temoporfin, temozolomide, teniposide, tesetaxel, testolactone, tetranitrate, Phosphinothioylidynetrisaziridine, tiazofurine, thioguanine, for pyrrole method, topotecan, ET-743, triaziquone, triethylene melamine, three Close platinum (Triplatin), retinoic acid, treosulfan, trofosfamide, uracil mustard, cut down soft than star, Verteporfin, vincaleucoblastine, Vincristine, vindesine, vinflunine, vinorelbine, Vorinostat, left soft than star and other Carbazole alkaloid described herein Agent or cytotoxic agent.
Owing to some medicines are used together more preferable than being used alone, therefore the most simultaneously with two or more drug administrations. Generally, use two or more chemotherapeutic agents as combinatorial chemistry therapy.In some embodiments, chemotherapeutic agent Agent (including combinatorial chemistry therapy) can be applied in combination with compound described herein.
Targeted therapies
In some embodiments, compound described herein is to use together with one or more targeted therapies.Targeting Therapy constitutes the cancer cell proteins matter to imbalance and has the use of specific medicament.Some little targeted molecular therapy medicines lead to Be often suddenly change in cancerous cell, process LAN or the inhibitor of enzymatic domain on other key proteins.Highlight Example is tyrosine kinase inhibitor, and such as Axitinib, bosutinib, AZD2171, Dasatinib, Erlotinib, she replaces by horse Buddhist nun, gefitinib, Lapatinib, lestaurtinib, Buddhist nun strangle for Buddhist nun, SU5416, Sorafenib, Sutent and Fan Deta Buddhist nun, and cell cycle protein dependent kinase inhibitor, such as Avobenzene former times ground (Alvocidib) and Sai Lixibu (Seliciclib).Monoclonal antibody therapy is another kind of strategy, and in this therapy, therapeutic agent is that specific bond is in cancerous cell table The antibody on protein on face.Example includes the anti-HER2/neu antibody trastuzumab being typically used in breast carcinomaAnd the anti-CD 20 antibodies Rituximab being typically used in multiple B cell malignant disease and Tositumomab.Other exemplary antibodies includes Cetuximab, Victibix, Herceptin, Ah coming organizes monoclonal antibody, shellfish cuts down list Anti-, edrecolomab and gemtuzumab Ozogamicin Mylotarg CDP 771.Exemplary fusion proteins includes VEGF Trap and denileukin.In some embodiments In, targeted therapies can be applied in combination with compound described herein.
Exemplary other therapeutic agent can also include EGF-R ELISA (EGFR) inhibitor, and the most western appropriate former times is single Anti-, Victibix, gefitinib, Erlotinib, Buddhist nun's trastuzumab, horse trastuzumab, prick calamite monoclonal antibody (zalutumumab) or Lapatinib.Resistance for EGFR inhibitor can become mesenchyme phenotype or mesenchyme phenotype to occur because of cells switch, and Have EGFR sudden change and mesenchyme phenotype tumor can for EGFR inhibitor have lower sensitivity (see for example, Sequist etc., (2011) Sci Transl Med.3:75；Buck etc., (2007) Mol Cancer Ther.6:532； Thomson etc., (2008) Clin Exp Metastasis 25:843).
Exemplary other therapeutic agent may also include glutathione depletors, and such as L-butylthiamine acid-(S, R)-sulfoxide is sub- Amine (BSO).
Exemplary other therapeutic agent can also include phosphoinositide 3-kinase (PI3K) inhibitor, such as perifosine, De Laxibu (Idelalisib), BKM120, PX-866, IPI-145, NVP-BEZ235, GDC0941 and BAY 80-6946.
Exemplary other therapeutic agent may also include heat shock protein 90 (HSP90) inhibitor, such as geldanamycin, root Red shell rhzomorph, 17-N-allyl amino-17-AAG (17AAG), lattice Buddhist nun are for cloth (ganetespib), 4- (4-(2,3-dihydro-1,4-benzo dioxine-6-base)-5-methyl isophthalic acid H-pyrazole-3-yl)-6-ethyl resorcinol, AUY922 (NVP-AUY922), BIIB021, STA9090, AT13387, NVP-BEP800 and SNX-2112 (PF-04928473).
Targeted therapies may also refer to the little peptide as " target-seeking equipment (homing device) ", and they can be combined in swollen Cell surface receptor around tumor or affected extracellular matrix.If connected to the radioactive nucleus of these peptides (such as, RGD) Element decays near cancerous cell, then this nucleic kills this cell the most at last.The example of this therapy includes
Immunotherapy
In some embodiments, compound described herein is to use together with one or more immunotherapies.Cancer Immunotherapy refers to that the immune system being designed for inducing patient self is to one group of various therapeutic strategy of antineoplastic.For producing Current methods for the immunne response of tumor includes BCG immunotherapy in the capsule of superficial bladder carcinoma, and uses interference Element and other cytokine induce immunoreation in renal cell carcinoma and melanoma patients.
Allogeneic Hematopoietic Stem Cell Transplantation is regarded as a kind of form of immunotherapy, because the immunocyte of donor is frequent Tumor will be attacked with Graft-versus-tumor response.In some embodiments, immunotherapy agents can be with described hereinization Compound is applied in combination.
Hormonotherapy
In some embodiments, compound described herein is to use together with one or more hormonotherapies.Some The growth of cancer can suppress by providing or blocking some hormone.The Common examples of hormone-sensitive tumor includes some class The breast carcinoma of type and carcinoma of prostate.Removal or blocking-up estrogen or testosterone are often important additional treatment.In some cancer, With Hormone agonists such as progestin administration may treatment on useful.In some embodiments, hormonotherapy medicament can be with Compound described herein is applied in combination.
Nutrient limitation diet
In some embodiments, compound described herein is to be combined with one or more nutrient limitation diet to execute With.Owing to cancerous cell relies on glucose to produce cellular energy, therefore reduce Fructus Vitis viniferae by carbohydrate restriction and protein Sugar blood level can suppress the growth of certain cancers.In some cancer, nutrient limitation diet, such as heat restriction, fasting And high fat diet, may be useful in treatment.In some embodiments, this nutrient limitation diet can with retouch herein The compound stated is applied in combination.
Neural disorder
Compound described herein or compositions can be used for treatment or prevent from (such as, being exposed to by neuronal tissue's damage Ischemia or anoxic event, wound or the nervous tissue of chronic neurodegenerative disease) Neuronal cell death that causes." neural Unit's disease " be the sacred disease relevant to glutamate excitotoxicity or disease, such as by the god of such as apoplexy or ischemic event The cerebral ischemia caused through event or anoxia.Can be neuroprotective is effectively provided, such as to prevent god with this compounds for treating Amount through unit's cell death.
Compositions and route of administration
Compositions set forth herein includes compound set forth herein (such as compound described herein), and other Therapeutic agent (if present), its amount is effective to realize the tune to disease or disease symptoms (include described herein those) Joint.
Term " pharmaceutically acceptable carrier or adjuvant " refers to execute to experimenter together with compound provided herein Carrier or adjuvant, and this carrier or adjuvant do not damage its pharmacological activity and when with the chemical combination that be enough to deliver therapeutic dose When the dosage of thing is used nontoxic.
Pharmaceutically acceptable carrier, adjuvant or the medium that can be used in pharmaceutical composition provided herein include but not It is limited to, ion-exchanger；Aluminium oxide；Aluminium stearate；Lecithin；Self-emulsifying drug delivery systems (SEDDS), such as d-alpha-tocopherol Cetomacrogol 1000 succinate；Surfactant in pharmaceutical dosage form, as TWEEN Series (Tweens) or other be similar to Polymeric delivery matrices；Serum albumin, such as human serum albumin；Buffer substance, such as phosphate；Glycine；Sorbic acid；Pyrusussuriensis Acid potassium；The partial glyceride mixture of saturated vegetable fatty acid；Water；Salt or electrolyte, as protamine sulfate, disodium hydrogen phosphate, Potassium hydrogen phosphate, sodium chloride, zinc salt, silica sol, magnesium trisilicate；Polyvinylpyrrolidone；Material based on cellulose；Poly- Ethylene glycol；Sodium carboxymethyl cellulose；Polyacrylate；Wax；Polyethylene-polyoxypropylene block polymer；Polyethylene Glycol and sheep Hair fat.Cyclodextrin, as α-, β-and gamma-cyclodextrin, or the derivant of chemical modification, such as hydroxyalkyl cyclodextrin, including 2-and 3- HP-β-CD, or other derivant dissolved, it is also possible to favorably for strengthening the chemical combination of multiple formula described herein The delivery of thing.
Pharmaceutical composition provided herein can with per os, parenteral, by suck spraying, partly, per rectum, per nasal, Buccal, transvaginal or use via the storage tank implanted, preferably by Orally administered or used by injection.Medicine provided herein Compositions can comprise nontoxic pharmaceutically acceptable carrier, adjuvant or the medium of any routine.In some cases, preparation PH value can be adjusted with pharmaceutically acceptable acid, alkali or buffer, deliver strengthening the compound prepared or its The stability of form.As used herein, this term parenteral includes subcutaneous, Intradermal, intravenous, intramuscular, intraarticular, tremulous pulse In, in intrasynovial, breastbone, in sheath, intralesional and intracranial injection or infusion techniques.
Pharmaceutical composition can be to be the form of sterile injectable prepared product, such as aqueous or the oiliness of sterile injectable Suspension.Dispersion or wetting agent that this suspension can be suitable for according to technology as known in the art, use (e.g., such as, are told Temperature 80) and suspending agent prepare.Sterile injectable prepared product can also is that at nontoxic parenteral acceptable diluent or molten Sterile injectable solution in agent or suspension, such as the solution 1, in 3-butanol.The acceptable matchmaker that can use It is situated between and solvent has mannitol, water, Ringer's mixture (Ringer ' s solution) and isotonic sodium chlorrde solution.Additionally, routinely Use aseptic fixed oil as solvent or suspension media.For this purpose, the fixedness of any gentleness can be used Oil, including monoglyceride or two glyceride of synthesis.Fatty acid (such as oleic acid and glyceride ester derivatives thereof) can be used on injection In preparation, natural pharmaceutically acceptable oil (such as olive oil or Oleum Ricini), especially their polyoxyethylated versions, also It is such.These oil solutions or suspension can also contain long-chain alcohol diluents or dispersant, or carboxymethyl cellulose or conventional Prepare pharmaceutically acceptable dosage form (as emulsion with or suspension) similar dispersant.Purpose for preparation, it is also possible to Use other the conventional surfactant being usually used in preparing pharmaceutically acceptable solid, liquid or other dosage form, such as tween Series or this dish series (Spans), and/or other similar emulsifying agent or bioavailability reinforcing agent.
Pharmaceutical composition provided herein can be any oral acceptable dosage form oral administration, this dosage form include but It is not limited to, capsule, tablet, emulsion and waterborne suspension, dispersion liquid and solution.In the case of the tablet Gong orally using, Conventional carrier includes lactose and corn starch.Typically it is also added with lubricant, such as magnesium stearate.For with capsule form per os Clothes are used, and the diluent being suitable for includes lactose and the corn starch being dried.When waterborne suspension and/or emulsion oral administration Time, can be suspended or dissolved in active component combining in the oil phase of emulsifying agent and/or suspending agent.If desired, may be used To add some sweeting agent and/or flavoring agent and/or coloring agent.
Pharmaceutical composition provided herein can also be used for the suppository form for rectal administration.These compositionss can be passed through Being mixed with the nonirritant excipient being suitable for by compound provided herein and prepare, this excipient is at room temperature solid, but It is liquid under rectal temperature, and therefore will melt in the rectum and discharge active component.These materials include but not limited to, Cocoa butter, Cera Flava and Polyethylene Glycol.
When desired treatment relates to the region or the organ that are easily accessible to by local application, medicine provided herein The local application of compositions is applicable.For being applied to skin partly, pharmaceutical composition should suspend with containing or dissolve The ointment being suitable for of the active component in carrier configures together.Carrier bag for the local application of compound provided herein Include but be not limited to, mineral oil, Albolene, white vaseline, propylene glycol, polyethylene glycol oxide polyoxypropylene compound, emulsifying Wax and water.As an alternative, this pharmaceutical composition can be suspended or dissolved in in the carrier of applicable emulsifying agent with containing Reactive compound be suitable for washing liquid or emulsifiable paste configure together.The carrier being suitable for includes but not limited to, mineral oil, anhydrosorbitol Sugar alcohol monostearate, polysorbate60, spermaceti ester type waxes, cetearyl alcohol, 2-octyl dodecanol, benzyl alcohol and water.Herein The pharmaceutical composition provided by rectal suppository formulation or can also be applied to lower intestinal tract with applicable enema preparation partly. In transdermal patch is also included within partly.
Pharmaceutical composition provided herein can be used by nasal aerosol or suction.These compositionss are according to medicine In formulation art prepared by widely-known technique, and can use benzyl alcohol or other anticorrosion being suitable in normal saline Agent, the enhancing absorption enhancer of bioavailability, fluorocarbon and/or other solubilizing agent as known in the art or dispersion Agent, is prepared as solution.
When compositions provided herein comprises compound and one or more other treatments of multiple formula described herein During the combination of agent or preventive, this compound existed should be single medication side with the dosage level of this other medicament In case between about the 1% to 100% of usual application dosage, between the most about 5% to 95%.This other medicament can be made A part for multiple dose scheme is used dividually with compound provided herein.As an alternative, these medicaments can be as list A part for one dosage type low temperature, is mixed together in single compositions with compound provided herein.
Compound described herein can such as pass through intravenous, intra-arterial, subcutaneous, intraperitoneal, intramuscular or subcutaneous injection, Or per os, buccal, per nasal, across mucosa, use with ophthalmically acceptable prepared product form partly, or used by suction, dosage range from Per kilogram of body weight about 0.5 is to about 100mg, alternately, dosage between every dose of 1mg and 1000mg, every 4 to 120 hours, or root Use according to the needs of certain drug.Methods herein covers uses the compound of effective dose or compound composition to realize Effect that is desired or that state.Typically, pharmaceutical composition provided herein will the most about 1 time to about 6 times, or alternative Ground, uses with continuous infusion.This using can be used as chronic or acute therapy.Can be combined to produce single with carrier material The amount of the active component of dosage form will change with the host treated and specific application pattern.Typical prepared product will contain about The reactive compound (w/w) of 5% to about 95%.As an alternative, these prepared products contain the active ingredient of about 20% to about 80% Thing.
May require than those lower or higher dosage set forth above.Concrete dosage for any particular patient Many factors is will depend upon which, including the activity of the particular compound used, age, body weight, general health disease with therapeutic scheme Suffer from, sex, diet, time of application, discharge rate, drug regimen, the order of severity of disease, illness or symptom and the course of disease, patient Tendentiousness to disease, illness or symptom, and the judgement for the treatment of physician.
After the illness of patient is improved, if necessary, it is possible to the dimension of compound provided herein, compositions or combination Hold dosage to use.Subsequently, with the change of symptom, can by applied dose or frequency or both, be reduced in these symptoms The level of the illness of this improvement is kept in time being relieved to desired level.But, patient may be required for disease symptoms Any recurrence carries out long-term intermittent treatment.
Patient selects and monitoring
Compound described herein can suppress transglutaminase.Therefore, can by firstly evaluate patient and/or experimenter with Determine that this experimenter, the need of suppression transglutaminase, selects patient and/or experimenter to use chemical combination described herein Thing is treated, if this experimenter is confirmed as needing glutamine enzyme level, then use described herein to this experimenter Compound.
Method as known in the art can be used, such as by measuring existence and/or the activity of patient's GLN enzyme, It is evaluated as experimenter needing glutamine enzyme level.In some embodiments, to the activity of cancer GLN enzyme and/ Or level is estimated.
Can such as the patient to receiving compound described herein that improves for illness and/or ill effect supervise Survey.The improvement of patient's illness can such as be commented by growth, the shortage of growth or disappear of monitoring cancer (such as tumor) Estimate.In some embodiments, use the assessment of radioactivity determination or hemolysis parameters that patient is estimated.
Patient and/or experimenter can be selected in the following manner to use compound described herein to treat: Optionally obtain Patient Sample A；Assess this sample to determine whether this sample is characterised by i) relatively low compared with reference standard E- Cadherin expression, ii) Vimentin level higher compared with reference standard, and/or iii) and reference standard Compare pyruvate carboxylase expression that is relatively low or that reduce；And if it is determined that this patient has compared with reference standard relatively low CAM 120/80 expression, or Vimentin level higher compared with reference standard, then with described hereinization Compound is used to this patient.
In some embodiments, by the expression of CAM 120/80 compared with reference standard, wherein this reference mark Standard is the expression of CAM 120/80 in epithelial cell, as any one in documents below is characterized: (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (supplementary issue 7): vii89；Thiery Deng, (2002) Nature Reviews Cancer 2 (6): 442).In some embodiments, compared with this reference standard, E- The expression of cadherin is relatively low, reduce or do not exist.In some embodiments, the expression of CAM 120/80 is logical The level crossing the RNA to coding CAM 120/80 is estimated measuring.In some embodiments, the expression of CAM 120/80 Level is that the protein expression level by CAM 120/80 is assessed.In some embodiments, the expression of CAM 120/80 Level is lower than this reference standard by least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.In some embodiments, the expression of CAM 120/80 has at least compared with this reference standard in terms of expression 1.5, the reduction of 2,5,10,15,20,25,50,75,100 times.
In some embodiments, by the expression of Vimentin compared with reference standard, wherein this reference standard It is Vimentin expression in epithelial cell, as below with reference in document, any one characterizes: (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (suppl 7): vii89； Thiery etc., (2002) Nature Reviews Cancer 2 (6): 442).In some embodiments, the table of Vimentin The level of reaching is by being estimated measuring to the level of the RNA of coding waveforms albumen.In some embodiments, Vimentin Expression be that the protein expression level by Vimentin is assessed.In some embodiments, the table of Vimentin The level that reaches is higher than this reference standard by least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80% Or 90%.In some embodiments, the expression of Vimentin has at least compared with this reference standard in terms of expression 1.5, the increase of 2,5,10,15,20,25,50,75,100 times.
In some embodiments, the expression of pyruvate carboxylase is relatively low or reduction compared with reference standard, wherein This reference standard is pyruvate carboxylase expression in epithelial cell, as below with reference in document, any one characterizes : (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (suppl 7):vii89；Thiery etc., (2002) Nature Reviews Cancer 2 (6): 442).In some embodiments, acetone The expression of acid carboxylase is higher or increase compared with this reference standard.In some embodiments, pyruvate carboxylase Expression is by being estimated measuring to the level of the RNA of encoding pyruvate carboxylase.In some embodiments, third The expression of keto acid carboxylase is that the protein expression level by pyruvate carboxylase is assessed.In some embodiments In, the expression of pyruvate carboxylase is higher than this reference standard by least 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 80% or 90%.In some embodiments, the expression of pyruvate carboxylase and this reference standard Compare the increase having at least 1.5,2,5,10,15,20,25,50,75,100 times in terms of expression.
Patient Sample A
Term " Patient Sample A ", " Samples subjects " with " sample " used interchangeably herein.Patient Sample A can be group Knit, or body fluid, or health product.Tissue sample can include that fix, paraffin-embedded, the fresh or sample of freezing.Citing For, tissue sample can include biopsy, cheek swab.Example organization include lung, breast, brain, nervous tissue, kidney, Ovary, thyroid, pancreas, colon, prostate, lymph node, skin, hair follicle and fingernail.Exemplary sample includes deriving from entity The sample of tumor.Exemplary body fluid includes blood, blood plasma, urine, lymph fluid, tear, perspiration, saliva, seminal fluid and cerebrospinal fluid. Exemplary health product includes expired gas.
This tissue, fluid or product can take out from patient and be analyzed.Assessment can include following one or more: Carry out the analysis to this tissue, fluid or product；Require the analysis of this tissue, fluid or product；Require from this tissue, fluid or The result that the analysis of product obtains；Or receive the result that the analysis from this tissue, fluid or product obtains.
Can be for the expression of gene described herein (such as, CAM 120/80, Vimentin, pyruvate carboxylase) This sample tissue, fluid or product are analyzed.Can be for protein described herein (such as, CAM 120/80, waveform Albumen, pyruvate carboxylase) expression this sample tissue, fluid or product are analyzed.Can be for the letter of preliminary election (such as, epithelium is to mesenchyme transition pathway, CAM 120/80 approach, Vimentin approach or acetone for number approach or phenotype approach Acid carboxylase approach) a kind of gene or the gene expression dose of several genes this sample tissue, fluid or product are carried out Analyze further.Can (such as epithelium be to mesenchyme transition pathway, CAM 120/80 for the signal pathway of preliminary election or phenotype approach Approach, Vimentin approach or carboxylase enzymatic pathway) a kind of protein or the protein expression level of multiple proteins This sample tissue, fluid or product are further analyzed.
The method of assessment sample
The expression of gene described herein (such as, CAM 120/80, Vimentin and pyruvate carboxylase) can make Method with the multiple well-known expression transcribing molecule, gene, protein, mRNA, genomic DNA or cDNA for detection In any one be evaluated.Gene expression can pass through the measurement of genetic transcription thing (such as, mRNA), by the albumen of translation The amount of matter measure or measure by the measurement of gene product activity or monitor；Any of which all can use this area Standard technique known to the skilled person is measured.The limiting examples of these methods includes nucleic acid hybridization, nucleic acid reverse transcription Method, nucleic acid amplification, immunization method, protein purification method, protein function or activation measurement for protein detection.
CAM 120/80
CAM 120/80 gene is positioned on human chromosome 16.CAM 120/80 is that the classical calcium of cadherin superfamily glues egg In vain.Coded CAM 120/80 is Ca-dependent cell-cell adherence glycoprotein, comprises five extracellular Ca2＋ mucin weights Again, cross-film district and the cytoplasmic tail of high conservative.Sudden change in this gene with include gastric cancer, breast carcinoma, colorectal carcinoma, Thyroid carcinoma and ovarian cancer are correlated with in interior cancer.Expection CAM 120/80 function forfeiture by increase propagation, invasion and attack and/or Shift and contribute to cancer progression.The ectodomain of this protein has mediated the antibacterial adhesion to mammalian cell, cell Matter domain is required by internalization.The CAM 120/80 transcript variant identified is drawn by the sudden change of consistent splice site Rise.
Vimentin
Vimentin dna is positioned on human chromosome 10, the member of coding intermediate filament protein family.Median fiber, even Together form the cytoskeleton of cell with micro-pipe and actin filament, it contributes to maintaining cell shape and cytoplasmic complete Whole property, and stablize cytoskeleton interaction.Vimentin also acts as mediated immunity response, controls low density lipoprotein, LDL source Cholesterol transport the effect of esteratic site from lysosome, and act as adhering to, migrating and involved in cell signalling And the organizer of multiple key proteins.
Pyruvate carboxylase (PC)
PC gene is positioned on human chromosome 11, coded protein pyruvate carboxylase, and this enzyme catalysis carboxylase becomes grass Ethyl acetoacetic acid.This organized enzyme is to be arranged in the tetrahedral homotype tetramer, and it is only located in mitochondrial matrix.Pyruvate carboxylase is joined With multiple cell processes, generate including glyconeogenesis, fat, insulin secretion and the synthesis of neurotransmitter glutamate.In this gene Sudden change is associated with pyruvate carboxylase deficiency.Identify and there are different 5 ' UTR but encoded replacing of this same protein Transcript variant for property montage.
Nucleic acid molecules
Method described herein can relate to based on corresponding to gene described herein (such as CAM 120/80, Vimentin, Pyruvate carboxylase) the nucleic acid of separation, the mRNA level in-site of such as CAM 120/80, the mRNA level in-site of Vimentin, acetone acid The mRNA level in-site of carboxylase, the expression of gene described herein to sample evaluating.As used herein, term " nucleic acid " or " nucleic acid Molecule " it is intended to include DNA molecular (such as, cDNA or genomic DNA) and RNA molecule (such as, mRNA) and use nucleotide The analog of DNA or RNA that analog produces.This nucleic acid molecules can be strand or double-strand.
" separation " nucleic acid molecules is the separate core of other nucleic acid molecules present in the natural origin with this nucleic acid molecules Acid molecule." separation " nucleic acid molecules can be free of and adjoins this core natively in obtaining the genomic DNA of organism of this nucleic acid The sequence (such as the sequence of coded protein) of acid (that is, being positioned at the sequence at 5 ' and 3 ' ends of this nucleic acid)." separation " nucleic acid divides Son, such as mRNA, can be substantially free of other cell material in the cell or tissue source obtaining this nucleic acid or other pollutes Property protein.
Nucleic acid molecules described herein can use standard molecular biological technique and well known by persons skilled in the art count In recording according to storehouse, available sequence information separates.Use all or part of of these nucleotide sequences, standard can be used miscellaneous Hand over and clone technology (such as, compile such as Sambrook etc., Molecular Cloning:A Laboratory Manua, second edition, Cold Spring Harbor Laboratory Press, described in Cold Spring Harbor, NY, 1989) separate this The nucleic acid molecules that literary composition describes.
Nucleic acid molecules described herein can use cDNA, mRNA or genomic DNA conduct according to standard PCR amplification technology Template and suitable oligonucleotide primers expand.The nucleic acid molecules thus expanded can be cloned in suitable carrier, and Characterized by DNA sequence analysis.Additionally, all or part of oligonucleotide corresponding to nucleic acid molecules can pass through standard Synthetic technology, prepares for example with automatization's DNA synthesizer.
The nucleic acid molecules separated can comprise such nucleic acid molecules, and it has and the nucleic acid corresponding to gene described herein Nucleotide sequence, or with coding corresponding to the nucleoside of nucleotide sequence complementary of the nucleic acid of the protein of gene described herein Acid sequence.Substantially complementary with the nucleotide sequence that the nucleic acid molecules of given nucleotide sequence complementary is given with this make it The nucleic acid molecules of stable duplex can be consequently formed with this given nucleotide sequence hybridization.
Nucleic acid molecules described herein can only comprise a part for nucleotide sequence.These nucleic acid molecules can be used for example as probe Or primer.Probe/primer can be the oligonucleotide of one or more generally purification.Sequence based on nucleic acid molecules described herein The probe of row can be used for detecting the transcript corresponding to gene described herein or genome sequence.This probe can comprise labelling base Group, such as radiosiotope, fluorescent chemicals, enzyme or enzyme cofactor.These probes can be used as the one of diagnostic test reagent box Part, this test kit is as by surveying the level of the nucleic acid molecules encoding this protein in the cell sample of patient Amount, such as detection mRNA level in-site identifies the cell or tissue expressing this protein.
The method of detection gene expression
The method of detection and/or quantitatively genetic transcription thing (such as mRNA or the cDNA being generated by) may include but be not limited to Southern blotting technique analysis, rna blot analysis, polymerase chain reaction (PCR) are analyzed and probe assay.Detection and/or quantitatively genetic transcription The method of thing (such as mRNA or the cDNA being generated by) may include but be not limited to method based on hybridization, such as with to this gene The hybridization that transcript (such as mRNA or the cDNA being generated by) special probe is carried out.Genetic transcription thing (such as mRNA or by Its cDNA produced) level can be by by this sample, or this mRNA or the cDNA that is generated by, or the sample expanded by it It is applied to nucleic acid microarray or chip array is measured.
The level of genetic transcription thing (such as mRNA or the cDNA being generated by) can be by based on polymerase chain reaction (PCR) Method, such as quantitative PCR, quantitatively in real time PCR, real-time PCR, reverse transcription PCR, Real time RT-PCR are measured.Gene The level of transcript (such as mRNA or the cDNA being generated by) can be by method based on order-checking, and such as quantitatively RNA order-checking comes It is measured.
The level of genetic transcription thing (such as mRNA) can be determined by original position as known in the art or in vitro method.Right In in vitro method, the available separation being not directed to mRNA and any RNA isolation technics of selecting are from sample, such as from sample Cell purification RNA (see, e.g., Ausubel etc. to compile, Current Protocols in Molecular Biology, John Wiley&Sons,New York 1987-1999).It addition, technology well known to the skilled person can be used, as such as The single stage RNA separating technology of bur Xin Siji (Chomczynski) (1989, United States Patent (USP) No.4,843,155) is the most right A large amount of tissue samples are treated.For in-situ method, mRNA is before testing without separating from cell.These sides In method, known Histological method can be used to prepare/process cell or tissue sample.This sample can be fixed on holder subsequently On, then contacting with probe, this probe can be with the mRNA hybridization of coding genes of interest transcript.
Mensuration can absolute expression levels based on genetic transcription thing (such as mRNA), normalized expression or relative table Reach level.Can by by the expression of genetic transcription thing and the another kind of gene stably expressed (such as constitutive expression House-keeping gene) expression compare, its absolute expression levels is corrected, by expression normalization.For returning One gene being suitable for changed includes house-keeping gene, such as histone H3 gene or actin gene.This normalization allows one Individual sample and another sample, the first sample such as obtained from patient with from this same patient (such as from another tissue or When different time points) the second sample of obtaining；Or between the sample of separate sources, such as from the trouble of a patient Person's sample compares with the expression of the Patient Sample A from another one patient.
The expression provided can be as relative expression levels.This relative expression levels can be by by this genetic transcription thing The absolute expression levels of (such as mRNA) relatively determines compared with reference standard.This reference standard may be included in genotype or phenotype The expression of the genes of interest transcript in the sample determined.This reference standard can be to be epithelium in genotype or Phenotypic characterization The expression water of the genes of interest transcript (such as, CAM 120/80, Vimentin, pyruvate carboxylase) in the cell of cell Flat.Epithelial cell can be as characterized in the following documents any one: (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (suppl 7): vii89；Thiery etc., (2002) Nature Reviews Cancer 2(6):442)。
Can at least two point in time measurement genetic transcription described herein things (such as, CAM 120/80, Vimentin, Pyruvate carboxylase) expression to determine whether there occurs the change of expression.For example, can be with described herein Compounds for treating before and after, or survey at one or more time points during treating by compound described herein Amount expression.If it find that this expression reduce, the expression of such as CAM 120/80 reduce compared with reference standard and/or The expression of Vimentin increase compared with reference standard, then can give with controlling that compound described herein is carried out to experimenter Treat.This reference standard can be by the expression of the genes of interest transcript in the epithelial cell characterized.Epithelial cell can lead to Cross method as known in the art, such as, as characterized in the following documents any one: (Yauch etc., (2005) Clin Cancer Res 11:24；Savagner etc., (2010) Ann Oncol.21 (suppl 7): vii89；Thiery etc., (2002) Nature Reviews Cancer 2(6):442)。
Protein
Method described herein can relate to based on corresponding to gene described herein (such as CAM 120/80, Vimentin, Pyruvate carboxylase) the protein of separation, the protein level of such as CAM 120/80, the protein level of Vimentin, The protein level of pyruvate carboxylase, the expression of gene described herein to sample evaluating.This may also comprise it biological alive Property part, variant, hypotype or the assessment of splice variant.Can use known to those skilled in the art by suitable purification schemes Standard protein purification technique, from sample, separate the natural polypeptides corresponding to destination protein matter.
" separation " or " purification " protein or its biologically-active moiety are substantially free of the cell obtaining this protein Or the cellular material in tissue-derived or other contaminative protein.Term " generally without cellular material " includes protein With the cellular component of the cell therefrom isolating this protein prepared by separate protein.The biologically-active moiety of polypeptide can include Comprise the aminoacid sequence of aminoacid sequence that is the most consistent with the aminoacid sequence of this protein or that derive from this protein Polypeptide, these polypeptide include the aminoacid more less than full length protein, and show corresponding full length protein at least one Activity.Typically, biologically-active moiety comprises domain or the motif of at least one activity with respective egg white matter.
The method of detection protein expression
The expression of protein or polypeptide can arbitrary by multiple means well known to the skilled person Plant and carry out detection with quantitative.Detection and/or quantitative protein described herein or polypeptide (such as, CAM 120/80, waveform egg In vain, pyruvate carboxylase) method may include but be not limited to, biochemical method, such as electrophoresis, capillary electrophoresis, efficient liquid phase Chromatography (HPLC), thin layer chromatography (TLC), super diffusion chromatography etc.；Or different immunoassays, such as fluid or gel Precipitation, immunodiffusion (unidirectional or two-way), immunoelectrophoresis, radioimmunoassay (RIA), enzyme-linked immunosorbent assay (ELISA), the cell sorting of immunofluorescence assay, western blot, immunohistochemistry, in situ hybridization, fluorescence-activation (FACS) etc..Technical staff can easily use known protein/antibody detection method to be used for determining whether cell expresses Protein described herein or polypeptide.
Protein or polypeptide can use immunoassay to detect.As used herein, immunoassay include utilizing specificity It is incorporated into the mensuration of the antibody of protein or polypeptide.Immunoassay can be tied by the specificity of detection protein or polypeptide and antibody Incompatible sign, from use other physically or chemically to separate, targeting and quantitatively this polypeptide different.This polypeptide can use multiple Generally acknowledge immunity combine measure any one detect and/or quantitatively (see, e.g., United States Patent (USP) No.4,366,241；4, 376,110；4,517,288；And 4,837,168).About the commentary of general immunoassay, referring further to Asai (1993) Methods In Cell Biology, volume 37: Antibodies in Cell Biology, Academic Press, Inc.New York；Stites and Terr (1991) Basic and Clinical Immunology, the 7th edition.For detecting and/or quantitative The immunoassay of protein or polypeptide can use various ways well known to the skilled person.
Can conjugated protein or the antibody of polypeptide, such as corresponding to protein described herein or polypeptide (such as, E-calcium Mucin, Vimentin, pyruvate carboxylase) the antibody with detectable label (labelling directly or indirectly), available In detecting this protein or polypeptide.Antibody can be polyclone or monoclonal.Complete antibody, or its fragment can be used, such as Fab or F (ab')2.For probe or antibody, term " labelling " is intended to by by detectable material and this probe Or antibody coupling (i.e., physically connect) and the probe that carries out or the direct labelling of antibody, and by with directly labelling Another kind of reagent reacting and the probe that carries out or the indirect labelling of antibody.The example of indirect labelling includes using fluorescently-labeled Two antibody test first antibodies, and with biotin, DNA probe is carried out end labelling so that it can use fluorescently-labeled chain Mould Avidin detects.This antibody can be also labelling (the most radiolabeled, chromophore label, fluorogen labelling or Enzyme labelling) antibody.Use antibody derivatives, such as with substrate or with protein-ligand to (such as biotin-strepto-is affine Element) protein or the antibody that combines of part, or the antibody height structure changes territory etc. of antibody fragment, such as single-chain antibody, separation, should Antibody derivatives specific bond in protein described herein, such as CAM 120/80, Vimentin, pyruvate carboxylase, as By corresponding to protein described herein or the gene of polypeptide (such as CAM 120/80, Vimentin, pyruvate carboxylase) Protein coded by the open reading frame of transcript, or experienced by its normal post translational modification all or part of this type of Protein or polypeptide.
Protein from cell can use technology well known to the skilled person to separate.The albumen used Matter separation method can for example, Harlow and Lane (Harlow and Lane, 1988, Antibodies:A Laboratory Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York) in retouched Those stated.
Expression can provide as relative expression levels.This relative expression levels can be by by the absolute table of this protein The level that reaches relatively determines compared with reference standard.Purpose egg in the sample that this reference standard may be included in genotype or phenotype determines The expression of white matter.This reference standard can be destination protein matter (example in genotype or cell that Phenotypic characterization is epithelial cell As, CAM 120/80, Vimentin, pyruvate carboxylase) expression.Epithelial cell can pass through side as known in the art Method characterizes, such as, as described in any one in documents below: (Yauch etc., (2005) Clin Cancer Res 11: 24；Savagner etc., (2010) Ann Oncol.21 (suppl 7): vii89；Thiery etc., (2002) Nature Reviews Cancer 2(6):442)。
Protein described herein or the expression water of polypeptide (such as, CAM 120/80, Vimentin, pyruvate carboxylase) Flat can at least at two point in time measurement to determine whether there occurs the change of expression.For example, can be with retouching herein Before and after the compound stated is treated, or one or more time during treating by compound described herein Point in time measurement expression.If it find that this expression decreases, the expression of such as CAM 120/80 and reference standard Compare reduction and/or expression increase compared with reference standard of Vimentin, then can give with described herein to experimenter The treatment that compound is carried out.
Test kit
There is also described herein test kit, these test kits include measuring gene described herein, and (such as, E-calcium glues egg In vain, Vimentin, pyruvate carboxylase) the means of gene expression dose.For example, this test kit can include can with this The examination that the gene expression product of the gene (such as, CAM 120/80, Vimentin, pyruvate carboxylase) that literary composition describes interacts Agent.This test kit can include multiple can be with several genes described herein (such as, CAM 120/80, Vimentin, acetone acid Carboxylase) gene expression product interact reagent.This reagent may include but be not limited to, antibody, Multiple Antibodies, oligonucleoside Sour or multiple oligonucleotide.This gene expression product may include but be not limited to, the molecule transcribed, RNA molecule, polypeptide, protein, Genomic DNA or cDNA.
This test kit the most optionally includes the reagent for carrying out mensuration described herein.For example, this test kit Can include buffer agent, solvent, stabilizer, preservative, purification column, detectable and enzyme, they are probably and separate from Patient Sample A Nucleic acid, such as, expand sample by qRT-PCR, and sample be applied to reagent described above, or from experimenter's sample Product separation protein and sample is applied to necessary to reagent described above；Or for Samples subjects directly should Reagent for reagent described above.Test kit may also include the positive and negative control sample, such as, compare nucleic acid samples (example As, from non-oncological patients or non-tumor tissue sample or the experimenter not accepting treatment of cancer or for experimenter Sample carries out the nucleic acid samples of other test sample tested simultaneously).Test kit may also include illustrative material, and this material can provide Relevant collect and process Patient Sample A, these samples are applied to the finger that gene expression dose measures and explains measurement result Lead.
The component of test kit can provide in any form, such as liquid, dry, semiarid or lyophilized form, or The form stored under lyophilisation condition.Typically, the component of test kit is to provide in sterile form.When reagent provides molten at liquid Time in liquid, this liquid solution is generally aqueous solution, such as aseptic aqueous solution.When reagent provides in a dry form, generally pass through Add the solvent being suitable for realize rebuilding.This solvent of such as sterile buffer optionally provides in this test kit.
This test kit can include a kind of or multiple container, for the examination of dress concentration be applicable to gene expression dose measures Agent box component, or with the description about the dilution in this mensuration.This test kit can comprise the list for these mensuration components Only container, allotter or compartment, and information material.For example, positive and negative control sample can be included in bottle or In bottle, the most compatible grader can be sealed in aseptic plastic packaging, and information material can be included in plastic bushing Or in bag.This test kit can include the container that multiple (such as, a bag) is single, each comprises one or more units of reagent Form (such as, for once measuring).The container of these test kits can be bubble-tight and/or waterproof.This container can be for Purposes is marked.
This test kit can include the information material for carrying out and explain this mensuration.This test kit may also provide relevant at what The guidance of the result of this mensuration is reported at place, such as, remove therapeutic community or health care provider.This test kit can include for reporting Accuse the form of gene activity measurement result described herein, and relevant these forms or other relevant information of whither sending Address and contact details, or the relevant URL (unified resource of report result in online database or application on site (such as, app) Localizer) address.In another embodiment, this information material can include the result about depending on this mensuration, and whether patient The guidance with anticancer stem cell agent therapy should be accepted.
The information material of these test kits is not limited to its form.In many cases, information material (such as description) It is to provide with the form of leaflet, such as printed text, picture and/or photo, such as labelling or printed sheet.But, provided Information material can be also other form, such as computer readable-material, records a video or records.The information material of this test kit can be contact Information, such as physical address, e-mail address, webpage or telephone number, wherein the user of this test kit can obtain correlation gene work Property measure and/or the substantive information of its application in method described herein.This information material can be with multiple format Any combination provides.
Samples subjects may be provided to measure supplier, such as ISP (such as the third-party institution) or medical services Supplier, it will be assessed sample in mensuration and provide reading result.For example, measure supplier and can receive sample from experimenter Product, such as tissue sample, or blood plasma, blood or blood serum sample, use described herein mensuration to assess this sample, and determine that this is subject to Examination person is to accept with the candidate of inhibitor for treating as described herein.This mensuration supplier may be notified that health care provider should Experimenter is the candidate with inhibitor for treating as described herein, and uses inhibitor described herein to this candidate. The result that this mensuration supplier can assess, and optionally about the one or many in diagnosis, prognosis or appropriate therapy selection Kind result with any applicable form, as by mail, or electronically, or be supplied to such as medical by online database ISP, or patient, or insurance company.The information collected by this mensuration supplier and provide can be stored in data base.
Embodiment
Embodiment A
In the present embodiment, measured the enzymatic activity of transglutaminase by the endpoint determination of coupling.By glutamine and Phosphate is each with equal to Km and AC50Concentration be supplied to GAC, and adjust GAC concentration to obtain linear response, continue 60 points Clock.Produced glutamate, Glu is changed into 2-OG by the glutamte dehydrogenase of kinetics excess.NAD is set to by this second step 2X Km, because the NAD of excess is inhibition.But, the third conjugate enzyme diaphorase of kinetics excess reclaims from NADH NAD is so that the concentration of NAD keeps constant during the time-histories of this mensuration.Diaphorase, also supplies with kinetics excess, it The NADH produced by GDH is aoxidized back NAD, is reduced into the resorufin of high fluorescence with "diazoresorcinol" simultaneously.Stopping being somebody's turn to do with SDS After mensuration, measure resorufin by Ex544/Em590.Reducing of signal indicates the suppression of certain component in coupled enzymatic system. Hit thing (hit) likely is screened (counterscreen) only for GDH/ diaphorase is counter, to remove second The hit thing of the coupled enzymatic system in mensuration.
1. material
2. buffer
2X buffer (300mM NaCl, 100mM HEPES pH 8.5,0.1%BSA, 0.5mM EDTA, 100mM phosphoric acid Sodium pH 8.5)
(1X buffer Final concentration, containing 13mM glutamine, 100 μMs of "diazoresorcinol"s, 50 μ g/ml cardiac muscles for 5X substrate mixture Yellow enzyme)
(1X buffer Final concentration, containing 0.875 μ g/ml GAC, 1.56mM NAD, 6.25 units/ml for 1.2X enzymatic mixture GDH)
Termination mix (ddH26%SDS in O)
Response procedures
1. add the 1 μ l 100%DMSO containing compound
2. add 40 μ l enzymatic mixtures and at room temperature hatch 60 minutes
3. add 10 μ l substrate mixture to start reaction
4. terminate reaction with the 6%SDS of 25 μ l and read Ex544Em 590
Embodiment B:
In the present embodiment, by the endpoint determination of coupling, the conjugate enzyme of compound suppression transglutaminase HTS method is surveyed The potentiality determining system are tested, and this system includes glutamte dehydrogenase and diaphorase.Glutamate, Glu is supplied to Km GDH, then carries out reducing deacylated tRNA amine to produce 2OG.NAD is fed to this system with 2X Km, and by the activity of diaphorase Monitor its conversion to NADH.Diaphorase is to be supplied to GDH with a large amount of kinetics excess, it NADH is converted back into NAD with Make NAD level keep constant in the reaction, resazurin reduction is become the resorufin of high fluorescence simultaneously.This survey is being terminated with SDS After Ding, measure resorufin by Ex544/Em590.Reducing of signal indicates the suppression of certain component in coupled enzymatic system.
3. material
4. buffer
2X buffer (300mM NaCl, 100mM HEPES pH 8.5,0.1%BSA, 0.5mM EDTA, 100mM phosphoric acid Salt pH 8.5)
2X substrate mixture (1X buffer Final concentration, 40 μMs of "diazoresorcinol"s, 1.8mM glutamic acid, 20 μ g/ml diaphorases)
10X NAD mixture (1X buffer Final concentration, 12.5mM NAD)
(1X buffer Final concentration, by for the most linear measured GDH enzyme for 2.5X enzymatic mixture；Such as, as retouched herein State 0.05 unit/ml to obtain the final concentration of 0.02 unit/ml).
Response procedures
1. add the 1 μ l 100%DMSO containing compound
2. add 20 μ l enzymatic mixtures and at room temperature hatch 60 minutes
3. add 5 μ l NAD mixture
4. add 25 μ l substrate mixture to start reaction
5. terminate reaction with the 6%SDS of 25 μ l and read Ex544Em 590
Such as pass through the mensuration described in embodiment, the energy of compound described herein suppression transglutaminase can be tested Power.For succinctly, the IC of test during the inhibitory activity of these compounds is expressed as the mensuration of embodiment A or embodiment B in table 150。 The data of exemplary compounds are shown below in table 2.As shown, " A " refers to IC50< the glutamine enzyme level of 100nM Agent." B " refers to IC50Glutamine enzyme inhibitor between 100nM and 500nM." C " refers to IC50Between 500nM and 1000nM Glutamine enzyme inhibitor." D " refers to IC50Glutamine enzyme inhibitor between 1 μM and 2 μMs." E " refers to IC50At 2 μMs and Glutamine enzyme inhibitor between 10 μMs." N/A " refers to wherein not obtain IC50Compound.
Table 2
Embodiment 1
6,6'-(hexamethylene-1,3-diyl) double (pyridazine-3-alcohol) (I-5)
Step A:N1, N3-dimethoxy-N1, N3-dimethyl cyclohexane-1,3-diformamide (I-2):
100mL SOCl by stirring2In hexamethylene-1, (20g, 0.12mol, 2:1 are cis: trans mixing for 3-dicarboxylic acids Thing) solution is heated to refluxing overnight.Concentrated, and with in the next step, it is not necessary to it is further purified.
At 0 DEG C, to CH2Cl2In N, O-dimethyl hydroxyl amine hydrochlorate (34g, 0.36mol) dropping DIPEA (45g, CH 0.36mol) and subsequently2Cl2In hexamethylene-1,3-dicarbapentaborane dichloro made above.Mixture is at room temperature stirred Mix overnight.With saturated NaHCO3Aqueous solution quencher.Organic layer is washed with saline, and through Na2SO4It is dried.Evaporated and passed through Purification by flash chromatography, to obtain title compound I-2 (13g, 43%).
1H NMR(400MHz,CDCl3) cis δ: 3.68 (s, 6H), 3.14 (s, 6H), 2.74 (t, J=11.4Hz, 2H), 1.94 1.64 (m, 5H), 1.55 1.31 (m, 3H). trans δ: 3.71 (s, 6H), 3.24-3.35 (m, 2H), 3.19 (s, 6H), 1.84 (t, J=5.8Hz, 2H), 1.67 (m, 6H).
Step B:1,1'-(hexamethylene-1,3-diyl) diethyl ketone (I-3):
At-60 DEG C, N1, the N3-dimethoxy-N1 in the THF of stirring, N3-dimethyl cyclohexane-1,3-bis-formyl Amine I-2 (5g, 0.03mol) dropping methyl-magnesium-bromide (3M, 30mL).Make mixture be warmed up to room temperature, and stir 3 hours.With full The NH of sum4Cl aqueous solution quencher, uses CH2Cl2Extraction.Separate and evaporate organic layer to provide title compound I-3 (3g, 90%).
1H NMR(400MHz,CDCl3) cis δ: 2.37 (tt, J=12.1,3.0Hz, 1H), 2.15 (s, 6H), 2.07 (d, J=12.1,1H), 1.98 1.91 (m, 2H), 1.43 1.32 (m, 3H), 1.28 1.21 (m, 3H). trans δ: 2.73 (m, 2H), 2.18 (s, 6H), 1.87 (t, J=5.8Hz, 2H), 1.68-1.78 (m, 4H), 1.48-1.57 (m, 2H).
Step C:2-(morpholino-4-) acetate (I-4):
10g 2-glyoxalic acid is added batch-wise to 100mL morpholine at 10 DEG C.The mixture of gained is stirred at 100 DEG C 5h.Then it is cooled to room temperature, then filters, filtrate is ground to obtain title compound I-4 with MTBE.
1H NMR(400MHz,DMSO-d6)δ:8.35(b,1H),3.79(b,4H),2.85(b,4H)。
Step D:6,6'-(hexamethylene-1,3-diyl) double (pyridazine-3-alcohol) (I-5):
20g 2-(morpholino-4-) acetate solution in 25mL MeOH adds 5g 1,1'-(-hexamethylene-1,3- Diyl) diethyl ketone.The solution making gained refluxes 12h under argon gas.It is slowly added 3.0mL hydrazine hydrate to the solution of gained.Make institute The mixture obtained is under argon gas in 70 DEG C of 6h that reflux；Then room temperature it is cooled to.Filter the precipitate occurred, wash with MeOH and do Dry, to provide 4g cis-6,6'-(hexamethylene-1,3-diyl) double (pyridazine-3-alcohol).By mother liquor concentrations, and pass through flash chromatography Purification, to obtain the 6 of 1.5g, 6'-(hexamethylene-1,3-diyl) double (pyridazine-3-alcohol) (trans: cis=1.5:1).
1Cis δ: 12.77 (s, the 2H) of H NMR (400MHz, DMSO-d6), 7.46 (d, J=9.8Hz, 2H), 6.83 (d, J =9.8Hz, 2H), 2.68 (dt, J=12.0,3.4Hz, 2H), 1.97 (d, J=13.2Hz, 1H), 1.91-1.85 (m, 3H), 1.54-1.48 (m, 2H), 1.34 (qd, J=13.2,3.4Hz, 1H). trans δ: 11.10 (b.s., 2H), 7.26 (d, J= 9.6Hz, 2H), 6.96 (d, J=9.6Hz, 2H), 3.10 (quintet, J=5.8Hz, 2H), 2.09 (t, J=5.8Hz, 2H), 1.82-1.74 (m, 4H), 1.59 (m, 2H) .LC-MS:m/z (M+H)=273.2.
Embodiment 2
N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-(pyridine-2- Base) acetamide) (20)
Step A:(1R, 3S) double (the 6-chlorine pyridazine-3-base) hexamethylene (I-6) of-1,3-
By stirring in 2mL POCl3In 6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-3-alcohol) (100mg, 0.37mmol) solution is heated to 80 DEG C 10 minutes.Then evaporated, and used frozen water quencher.Alkalize with 2N NaOH Water layer, and use CH2Cl2Extraction.Organic layer is washed with saline, and through Na2SO4It is dried.Evaporated, and with quick post (PE:EA =2:1 to 1:1) purification, to obtain desired product (30mg, 26%).
1H NMR(400MHz,CDCl3) δ: 7.45-7.54 (d, J=8.9Hz, 2H), 7.36-7.44 (d, J=8.6Hz, 2H), 3.24 (t, J=10.7Hz, 2H), 2.31 (d, J=12.6Hz, 1H), 2.14 (d, J=5.9Hz, 3H), 1.98-2.11 (q, J=12Hz, 1H), 1.67-1.75 (m, 3H).LC-MS:m/z (M+H)=309.0
Step B:N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-(pyrroles Pyridine-2-base) acetamide) (20)
Double (the 6-chlorine pyridazine-3-base) hexamethylene of (1R, 3S)-1,3-in the 1mL dioxane that will stir (50mg, 0.16mmol)、Cs2CO3(210mg,0.64mmol)、Pd2(dba)3Double diphenylphosphine-the 9,9-of (15mg, 0.016mmol), 4,5- Dimethyl xanthene (xantphos) (14mg, 0.024mmol) solution was in 100 DEG C of microwave heatings 1 hour.Cooling reaction mixing Thing, then filtration over celite pad, wash with dioxane.Evaporation solvent, by quick post and preparation HPLC purification subsequently Residue, to obtain desired product (15mg, 18%).
1H NMR(400MHz,CDCl3) δ: 8.69 (d, J=4.3Hz, 2H), 8.47 (d, J=9.1Hz, 2H), 7.73 (td, J=7.7,1.5Hz, 2H), 7.37 (d, J=9.1Hz, 2H), 7.33 (d, J=7.8Hz, 2H), 7.25-7.31 (m, 2H), 4.06 (s, 4H), 3.14 (t, J=7.2Hz, 2H), 2.26 (d, J=12.1Hz, 1H), 2.09 (d, J=8.9Hz, 3H), 1.84-2.01 (q, J=12Hz, 1H), 1.57-1.74 (m, 3H).LC-MS:m/z (M+H)=509.6.
N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-phenylacetyls Amine) (10)
Program is identical with the program of embodiment 2 step B.
1H NMR(400MHz,CDCl3) δ: 9.91 (s, 2H), 8.46 (d, J=9.4Hz, 2H), 7.37-7.39 (m, 6H), 7.31-7.34 (m, 4H), 7.28-7.31 (m, 2H), 3.96 (s, 4H), 3.09 (t, J=11.8Hz, 2H), 2.21 (d, J= 12.1Hz,1H),2.05(m,3H),1.87-1.97(m,1H),1.50-1.70(m,3H).LC-MS:m/z (M+H)=507.2
N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) dipropyl acidamide) (152)
Program is identical with the program of embodiment 2 step B.
1H NMR(400MHz,CDCl3) δ: 8.58 (d, J=8.9Hz, 2H), 7.53 (d, J=9.1Hz, 2H), 3.16 (m, 2H), 2.63 (q, J=7.3Hz, 4H), 2.27 (d, J=8.6Hz, 1H), 2.13 (m, 3H), 1.99 (m, 1H), 1.69 (m, 3H), (1.25-1.31 t, J=7.3Hz, 6H).LC-MS:m/z (M+H)=383.4
N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) two butyramides) (37)
Program is identical with the program of embodiment 2 step B.
1H NMR(400MHz,CDCl3) δ: 10.41 (s, 2H), 8.53 (d, J=9.2Hz, 2H), 7.42 (d, J=9.3Hz, 2H), 3.12 (m, 2H), 2.66 (t, J=7.5Hz, 4H), 2.30 (d, J=12.8Hz, 1H), 2.12 (m, 3H), 1.95 (q, J= 12.5Hz, 1H), 1.77 (sextet, J=7.4Hz, 4H), 1.69 (m, 3H), 1.00 (t, J=7.4Hz, 6H).LC-MS:m/z (M+H)=411.4
N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-methyl propionyl Amine) (98)
Program is identical with the program of embodiment 2 step B.
1H NMR(400MHz,CDCl3) δ: 9.64 (s, 2H), 8.50 (d, J=9.2Hz, 2H), 7.42 (d, J=9.2Hz, 2H), 3.14 (t, J=11.5Hz, 2H), 2.93 (septet, J=6.5Hz, 2H), 2.30 (d, J=12.4Hz, 1H), 2.12 (m, 3H), 1.96 (q, J=11.8Hz, 1H), 1.71 (m, 3H), 1.28 (d, J=6.5Hz, 12H).
Embodiment 3
N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) diacetayl amide) (99)
Step A:(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) diamino acid hexichol Methyl ester (176)
Program is identical with the program of embodiment 2.
1H NMR(400MHz,CDCl3) δ: 8.22 (d, J=8.9Hz, 4H), 7.36-7.46 (m, 10H), 5.26 (s, 4H), 3.14 (t, J=11.8Hz, 2H), 2.26 (d, J=14.8Hz, 1H), 2.10 (m, 3H), 1.89-2.05 (m, 1H), 1.56- 1.76(m,3H).LC-MS:m/z (M+H)=539.2
Step B:6,6'-((1R, 3S)-hexamethylene-1,3-diyl) two pyridazine-3-amine (I-7)
At room temperature, by double for 6'-((1R, 3S) hexamethylene-1,3 diyls) (pyridazine 6,3-diyl) diamino acid benzene methyl (20mg, 0.037mmol) and 10mg 20%Pd (OH)2/ carbon suspension in 2mL MeOH is at H2Lower stirring 1.5h.Make reaction Mixture filtration over celite pad, and wash with MeOH.Evaporation filtrate, and by preparation HPLC purification, obtain desired product (7mg)。
1H NMR(400MHz,DMSO-d6) δ: 7.19 (d, J=9.1Hz, 2H), 6.71 (d, J=9.1Hz, 2H), 6.11 (s, 4H), 2.85 (t, J=11.7Hz, 2H), 1.93 (d, J=10.2Hz, 1H), 1.86 (m, 3H), 1.71 (m, 2H), 1.39- 1.62(m,4H).LC-MS:m/z (M+H)=271.1
Step C:N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) diacetayl amide (99)
By 6,6'-((1R, 3S)-hexamethylene-1,3-diyl) two pyridazine-3-amine (40mg, 0.145mmol), acetic acid (0.05mL, 0.45mmol), HATU (171.1mg, 0.45mmol) and N-ethyl-N-iospropyl acrylate-2-amine (62mg, 0.48mmol) solution in N,N-dimethylformamide (5mL) is heated to 50 DEG C overnight.Pour the mixture into water (20mL) In, precipitate is collected by filtration.By preparation TLC (ethyl acetate: methanol=10:1) pure solid, obtain desired product (7mg)。
1H NMR(400MHz,DMSO-d6) δ: 11.03 (s, 2H), 8.24 (d, J=9.1Hz, 2H), 7.67 (d, J= 9.4Hz, 2H), 3.09 (t, J=11.8Hz, 2H), 2.13 (s, 6H), 2.21 (d, J=12.1Hz, 1H), 2.05 (m, 3H), 1.87-1.97(m,1H),1.50-1.70(m,3H).LC-MS:m/z (M+H)=355.2
N, N'-(6,6'-((1R, 3S)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-(3-(fluoroforms Epoxide) phenyl) acetamide (180)
Program is identical with the program of step C of embodiment 3.
1H NMR(400MHz,CDCl3) δ: 11.68 (s, 2H), 8.50 (d, J=9.4Hz, 2H), 7.25-7.39 (m, 8H), 7.04-7.14 (m, 2H), 4.17 (s, 4H), 3.07 (t, J=11.8Hz, 2H), 2.23 (d, J=12.6Hz, 1H), 2.04 (m, 3H),1.85(m,1H),1.43-1.68(m,3H).LC-MS:m/z (M+H)=675.2.
Embodiment 4
Trans-(raceme)-N, N'-(6,6'-(hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-(pyrroles Pyridine-2-base) acetamide) (14)
Step A:6,6'-(trans)-hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl) double (fluoroform sulphonates) (I-8)
At 0 DEG C, to the 6 of stirring, and 6'-(hexamethylene-1,3-diyl) double (pyridazine-3-alcohol) (400mg, 1.5mmol, instead Formula: cis=1.5:1) at 5mL CH2Cl2In solution add DIPEA (570mg, 4.5mmol), then add Tf2O (846mg,3mmol).Make mixture be warmed up to room temperature, stir 3 hours.With saturated NaHCO3Aqueous solution quencher, uses CH2Cl2Extraction Take.Separate organic layer, wash organic layer with saline, and through Na2SO4It is dried.Concentrated, and by quick post (PE:EA=2:1 To 1:1) purification, to obtain desired trans product (350mg, 45%).
1H NMR(400MHz,CDCl3) δ: 7.71 (d, J=8.9Hz, 2H), 7.40 (d, J=8.9Hz, 2H), 3.62 (q, J =5.8Hz, 2H), 2.56 (t, J=5.8Hz, 2H), 2.03-2.14 (m, 4H), 1.74 (t, J=5.8Hz, 2H).LC-MS:m/z (M+H)=537.0
Step B: anti-form-1, double (the 6-bromine pyridazine-3-base) hexamethylene (I-9) of 3-
6,6'-(trans)-hexamethylene-1,3-diyl to stirring) double (pyridazine-6,3-diyl) double (fluoroform sulphonates) (350mg, 0.65mmol) is at 5mL CH3Solution in CN adds LiBr (280mg, 3.3mmol), be followed by HBr (0.32mL, 1.9mmol, in AcOH solution 33%).Heat the mixture to reflux 3 hours.Then concentrated, pure by quick post Change, to obtain the product (135mg, 52%) of white solid.
1H NMR(400MHz,CDCl3) δ: 7.62-7.74 (d, J=8.9Hz, 2H), 7.39-7.48 (d, J=8.9Hz, 2H), 3.56 (q, J=5.8Hz, 2H), 2.52 (t, J=5.8Hz, 2H), 1.98-2.17 (m, 4H), 1.65-1.82 (m, 2H). LC-MS:m/z (M+H)=397.0
Step C: trans-(raceme)-N, N'-(6,6'-(hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) is double (2-(pyridine-2-base) acetamide) (14)
By double for (1R, 3S)-1,3-(6-chlorine pyridazine-3-base) hexamethylene (40mg, 0.1mmol), Cs2CO3(98mg, 0.3mmol)、Pd2(dba)3Double diphenylphosphine-9,9-dimethyl the xanthene of (14mg, 0.015mmol) and 4,5-(12mg, 0.02mmol) mixture in 2mL dioxane was in 110 DEG C of microwave heatings 1.5 hours.Make the mixture filtration over celite of cooling Pad, and wash with dioxane.Then concentrate eluant, by preparation HPLC purification, to provide desired product (9mg).
1H NMR(400MHz,CDCl3) δ: 8.70 (d, J=4.4Hz, 2H), 8.41 (d, J=9.2Hz, 2H), 7.73 (td, J=7.6,1.2Hz, 2H), 7.43 (d, J=9.2Hz, 2H), 7.27-7.34 (m, 4H), 4.03 (s, 4H), 3.39 (quintet, J =5.6Hz, 2H), 2.44 (t, J=5.6Hz, 2H), 2.05 (m, 2H), 1.87-1.90 (m, 4H), 1.70 (m, 2H).LC-MS: M/z (M+H)=509.6
Trans-(raceme)-N, N'-(6,6'-(hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) diacetayl amide (41)
Program is identical with the program of step C of embodiment 4.
1H NMR (400MHz, DMSO-d6) δ: 8.19-8.26 (d, J=9.4Hz, 2H), 7.62-7.71 (d, J= 9.4Hz, 2H), 3.23-3.31 (quintet, J=5.8Hz, 2H), 2.31 (t, J=5.8Hz, 2H), 2.15 (s, 6H), 1.99 (m, 2H), 1.83-1.90 (m, 2H), 1.64 (d, J=5.9Hz, 2H).LC-MS:m/z (M+H)=355.5
Trans-(raceme)-N, N'-(6,6'-(hexamethylene-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-(4- (trifluoromethoxy) phenyl) acetamide) (75)
Program is identical with the program of step C of embodiment 4.
1H NMR(400MHz,CDCl3) δ: 8.50 (d, J=8.1Hz, 2H), 7.34-7.48 (m, 6H), 7.18 (d, J= 8.1Hz, 4H), 4.03 (s, 4H), 3.36 (quintet, J=5.8Hz, 2H), 2.38 (t, J=5.8Hz, 2H), 1.95-2.00 (m,2H),1.83-1.91(m,2H),1.61-1.65(m,2H).LC-MS:m/z (M+H)=675.6
Embodiment 5
2-phenyl-N-(6-((1S, 3R)-3-(5-(2-phenylacetamido)-1,3,4-thiadiazoles-2-base) cyclohexyl) Pyridazine-3-base) acetamide (291)
Step A: trans-(raceme)-3-(5-amido-1,3,4-thiadiazoles-2-base) cyclohexanecarboxylate (I-10)
By trans-3-(methoxycarbonyl) naphthenic acid (30g, 0.161mol) and thiosemicarbazides (18.2g, 0.2mol) At POCl3(80mL) solution in is heated to 40 DEG C and continues 30min, then 60 DEG C of lasting 30min, and then 80 DEG C continue 3h.When Consumption of raw materials is complete, enriched mixture, and residue 4N NaOH is neutralized to pH=8, then extracts by ethyl acetate (3 × 150ml) Mixture.It is dried the organic layer of merging with anhydrous sodium sulfate, filters, concentrate, by the crude product of flash column gained to obtain To desired product (12g, 30%).
1H NMR (400MHz, DMSO-d6) δ: 7.04 (s, 2H), 3.63 (s, 3H), 3.12 (dt, J=8.3,4.1Hz, 1H),2.68-2.83(m,1H),2.00-2.15(m,1H),1.71-1.94(m,3H),1.44-1.68(m,4H),LC-MS:m/z (M+H)=242.5
Step B: trans-(raceme)-3-(5-(2-phenylacetamido) 1,3,4-thiadiazoles 2-yl) cyclohexane-carboxylic acid Methyl ester (I-11)
At 0 DEG C, to trans-3-(5-amido-1,3,4-thiadiazoles-2-base) cyclohexanecarboxylate (6g, 0.024mol) at CH2Cl2(100mL) solution in adds triethylamine (4.85g, 0.048mol), is then slowly added 2-phenyl Chloroacetic chloride (4.6g, 0.029mol).Stirring mixture 4h.Then pour the mixture in 100mL water, and extract by ethyl acetate Take.It is dried organic layer with anhydrous sodium sulfate, filters, then concentrate to obtain yellow solid.This solid is ground by ethyl acetate, and Filter, to obtain title compound (8.5g, 0.023mol).LC-MS:m/z (M+H)=360.5
Step C: trans-(raceme)-N-methoxy-. N-methyl-3-(5-(2-phenylacetamido) 1,3,4-thiadiazoles 2 bases) cyclohexane carboxamide (I-12)
To trans-3-(5-(2-phenylacetamido)-1,3,4-thiadiazoles-2-base) cyclohexanecarboxylate (8.5g, 0.023mol) at CH3OH/THF (V:V=3:1；Solution in 120mL) adds containing LiOH H2O (1.98g, 0.047mol) Water (20ml).Stirring reactant mixture 3h, is concentrated in vacuo, and is extracted with ethyl acetate.Water layer 1M HCl is adjusted to pH =6.Filter the precipitate being consequently formed and be dried, to obtain trans-3-(5-(2-phenylacetamido)-1,3,4-thiadiazoles 2-yl) naphthenic acid (6g, 0.017mol).
LC-MS:m/z (M+H)=389.5
By trans-3-(5-(2-phenylacetamido) 1,3,4-thiadiazoles 2 base) naphthenic acid (6g, 0.017mol), N, O-dimethyl hydroxyl amine hydrochlorate (3.37g, 0.034mol), HATU (7.26g, 0.019mol) and N-ethyl-N-iospropyl Acrylate-2-amine (6.87g, the 0.068mol) heated overnight at 50 DEG C of the solution in 100mL DMF.Mixture is cooled down, then Pour in 100mL water, then extract by ethyl acetate (3 × 100ml).The organic layer merged with saline washing, uses anhydrous slufuric acid Sodium is dried, filters, concentrates, and by purification by flash chromatography, to obtain desired product (4.5g, 0.012mol).
LC-MS:m/z (M+H)=389.5
Step D: trans-N-(5-((1S, 3S)-3-acetyl group cyclohexyl)-1,3,4-thiadiazoles-2-base)-2-phenyl second Amide (I-13)
At 0 DEG C, to trans-N-Methoxy-N-methyl-3-(5-(2-phenylacetamido) 1,3,4-thiadiazoles-2- Base) cyclohexane carboxamide (4.5g, 0.012mol) solution interpolation CH in anhydrous THF (100mL)3MgBr(3.0M； 7.75mL).Then reactant mixture is stirred 2h, and by adding saturated NH4Cl aqueous solution quencher.With ethyl acetate (2 × 100ml) extraction mixture；And with the organic layer of saline washing merging, be dried through anhydrous sodium sulfate, filter and concentrate, to obtain Oily residue.By this residue of purification by flash chromatography to obtain product (3.0g, 73%).
1H NMR(400MHz,DMSO-d6)δ:12.69(s,1H),7.13-7.42(m,5H),3.80(s,2H),3.01- 3.20 (m, 1H), 2.58 (m, 1H), 2.20 (d, J=12.6Hz, 1H), 2.13 (s, 3H), 2.02 (d, J=12.1Hz, 1H), 1.83-1.96(m,2H),1.26-1.55(m,4H).LC-MS:m/z (M+H)=344.5
Step E: cis N-(5-(3-(6-hydroxypyridazin-3-base) cyclohexyl) 1,3,4-thiadiazoles-2-base) 2-phenyl second Amide (I-14)
By 4-(carboxy-methylene) morpholine-4-(1.25g, 8.75mmol) and trans-N-(5 ((1S, 3S) 3-acetyl group Cyclohexyl)-1,3,4-thiadiazoles-2-base)-2-phenyl-acetamides (1.5g, 4.37mmol) is at 50mL CH3Solution in OH adds Heat 100 DEG C is overnight.Cooling mixture, then concentrates.Thus obtained crude product is directly used in next step.To being dissolved in 20mL This crude product in n-BuOH adds N2H4·H2O (2ml), and heat the mixture to 130 DEG C of 4h.Cooling mixture is the denseest Contracting, and by purification by flash chromatography, with obtain 6-((1S)-3-(5-amido-1,3,4-thiadiazoles-2-base) cyclohexyl) pyridazine- 3-alcohol (400mg crude product).
LC-MS:m/z (M+H)=278.5.
To 6-((1S)-3-(5-amino-1,3,4-thiadiazoles-2-base) cyclohexyl) pyridazine-3-alcohol (400mg crude product, 1.44mmol) at CH2Cl2In solution add triethylamine (291mg, 2.88mmol), be followed by 2-phenylacetyl chlorine (266mg, 1.73mmol).Reactant mixture is stirred at room temperature 1h.Enriched mixture, and by purification by flash chromatography residue to obtain To title compound.
1H NMR (400MHz, DMSO-d6) δ: 12.79 (br.s., 1H), 12.68 (br.s., 1H), 7.48 (d, J= 9.7Hz, 1H), 7.32-7.35 (m, 2H), 7.22-7.31 (m, 3H), 6.83 (d, J=9.7Hz, 1H), 3.83 (s, 2H), 3.15-3.26 (m, 1H), 2.74 (t, J=12.1Hz, 1H), 2.19 (d, J=12.6Hz, 1H), 2.03-2.11 (m, 1H), 1.78-1.97(m,2H),1.36-1.61(m,4H)；LC-MS:m/z (M+H)=396.5.
Step F: cis-N-(5-(3-(6-chlorine pyridazine-3-base) cyclohexyl)-1,3,4-thiadiazoles-2-base)-2-phenyl second Amide (I-15)
By N-(5-(3-(6-hydroxypyridazin-3-base) cyclohexyl)-1,3,4-thiadiazoles-2-base) 2-phenyl-acetamides- (260mg, 0.66mmol) is at POCl3(5mL) solution in is heated to 80 DEG C and continues 30min.Cooling mixture, then concentrates, And with saturated Na2CO3Aqueous solution regulates to pH=8.Mixture is extracted by ethyl acetate (2 × 50ml).Merge with saline washing Organic layer, be dried through anhydrous sodium sulfate, filter, concentrate, and by preparative TLC purification, to obtain title compound (60mg, 22%).
1H NMR (400MHz, DMSO-d6) δ: 12.67 (br.s., 1H), 7.86 (d, J=8.9Hz, 1H), 7.80 (d, J =9.1Hz, 1H), 7.30-7.37 (m, 4H), 7.27 (m, 1H), 3.80 (s, 2H), 3.27 (m, 1H), 3.15 (m, 1H), 2.27 (d, J=12.9Hz, 1H), 2.11 (m, 1H), 1.91-2.00 (m, 2H), 1.86 (q, J=12.9Hz, 1H), 1.50-1.60 (m, 3H).LC-MS:m/z (M+H)=414.5.
Step G:2-phenyl-N-(6-((1S, 3R)-3-(5-(2-phenylacetamido)-1,3,4-thiadiazoles-2-base) ring Hexyl) pyridazine-3-base) acetamide (291)
Program is identical with the program of step B of embodiment 2.
1H NMR(400MHz,CDCl3) δ: 8.60 (d, J=9.1Hz, 1H), 7.49 (d, J=9.1Hz, 1H), 7.39- 7.43(m,4H),7.31-7.37(m,6H),4.03(s,2H),3.96(s,2H),3.29(m,1H),3.11(m,1H),2.45 (d, J=13.7Hz, 1H), 2.26 (m, 1H), 2.02-2.09 (m, 2H), 1.90 (q, J=13.7Hz, 1H), 1.54-1.72 (m, 3H).LC-MS:m/z (M+H)=513.5
Embodiment 6
N, N'-(6,6'-(Pentamethylene .-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-(pyridine-2-base) acetamide) (175)
Step A:N1, N3-dimethoxy-N1, N3-dimethylcyclopentane-1,3-diformamide (I-16):
By Pentamethylene .-1 of stirring, 3-dicarboxylic acids (20g, 0.13mol, 1:1 are cis: trans) is at 100mL SOCl2In Solution is heated to refluxing overnight.Cooling mixture, concentrates, and with in the next step, it is not necessary to it is further purified.
At 0 DEG C, to CH2Cl2In N, O-dimethyl hydroxyl amine hydrochlorate (34g, 0.36mol) dropping DIPEA (45g, CH 0.36mol) and subsequently2Cl2In Pentamethylene .-1,3-dicarbapentaborane dichloro made above.Mixture is at room temperature stirred Mix overnight.Used saturated NaHCO3Aqueous solution processes, and then washs organic layer with saline, and through Na2SO4It is dried, and evaporation Solvent.By purification by flash chromatography residue to provide desired product (20g, 65%).
1H NMR(400MHz,CDCl3) cis δ: 3.68 (s, 6H), 3.17 (s, 6H), 3.10 (m, 2H), 2.10 (m, 1H), 1.95 (m, 3H), 1.83 (m, 2H). trans δ: 3.68 (s, 6H), 3.32 (m, 2H), 3.17 (s, 6H), 2.07 (t, J=8Hz, 2H),2.00(m,2H),1.79(m,2H).LC-MS:m/z (M+H)=245.2
Step B:1,1'-(Pentamethylene .-1,3-diyl) diethyl ketone (I-17)
At-60 DEG C, to stirring N1, N3-dimethoxy-N1, N3-dimethyl cyclohexane-1,3-diformamide (5g, 0.02mol) dropping methyl-magnesium-bromide of the solution in THF (3M, 20mL).Make mixture be warmed up to room temperature, and stir 3 hours. With saturated NH4Cl aqueous solution processes, and then uses CH2Cl2Extraction.Be dried and evaporate organic layer, with obtain desired product (3g, 93%).
1H NMR(400MHz,CDCl3) cis δ: 2.91 (m, 2H), 2.17 (s, 6H), 2.07 (m, 1H), 1.92 (m, 1H), 1.86-1.88 (m, 4H). trans δ: 3.00 (quintet, J=7.6Hz, 2H), 2.17 (s, 6H), 2.01 (t, J=7.8Hz, 2H),1.89(m,2H),1.75(m,2H)。
Step C:4,4'-(Pentamethylene .-1,3-diyl) double (2-morpholino-4-ketobutyric acid) (I-18)
To the 2-(morpholino-4-) acetate (15g) solution in MeOH (25mL) add 1,1'-(-Pentamethylene .-1, 3-diyl) diethyl ketone (3g).Reflux under argon gas 12h by the solution of gained.Then concentrated, with CH2Cl2Dissolve, and filter To obtain title compound.It can use without being further purified.
LC-MS:m/z (M+H)=441.2
Step D:6,6'-(Pentamethylene .-1,3-diyl) double (pyridazine-3-alcohol) (I-19):
At room temperature, to the 4 of stirring, 4'-(Pentamethylene .-1,3-diyl) double (2-morpholino-4-ketobutyric acid) (8.7g, 0.02mol) solution in n-BuOH (50mL) adds hydrazine hydrate (1.0mL).The mixture making gained refluxes at 140 DEG C 12h, then cools down and concentrates.By purification by flash chromatography residue with obtain desired compound (trans: cis=1: 1%).
Cis δ: 12.77 (s, the 2H) of 1H NMR (400MHz, DMSO-d6), 7.21 (d, J=9.8Hz, 2H), 6.95 (d, J =9.8Hz, 2H), 3.22 (m, 2H), 2.38 (dt, J=12.8,7.6Hz, 1H), 2.13 (m, 1H), 1.86-1.88 (m, 4H). Trans δ: 12.77 (br.s., 2H), 7.26 (d, J=9.6Hz, 2H), 6.96 (d, J=9.6Hz, 2H), 3.30 (quintet, J= 7.4Hz, 2H), 2.20 (t, J=8.0Hz, 2H), 2.15 (m, 2H), 1.85 (m, 2H).LC-MS:m/z (M+H)=259.2
Double (the 6-chlorine pyridazine-3-base) Pentamethylene. (I-20) of step E:1,3-
By the 6,6'-(Pentamethylene .-1,3-diyl) double (pyridazine-3-alcohol) (1g, 0.38mmol) of stirring at POCl3(5mL) In solution be heated to 80 DEG C continue 30 minutes.Cooling mixture, then concentrates, and treats with frozen water.With in 2N NaOH and water Layer, and use CH2Cl2Extraction.Organic layer is washed with saline, and through Na2SO4It is dried.Remove solvent, and via purification by flash chromatography Residue, to provide desired product (600mg, 50%).
1H NMR (400MHz, DMSO-d6) cis δ: 7.57 (d, J=8.9Hz, 2H), 7.48 (d, J=8.9Hz, 2H), 3.73 (m, 2H), 2.72 (dt, J=13.0,7.6Hz, 1H), 2.31-2.43 (m, 3H), 2.16 (m, 2H). trans δ: 7.47 (d, J=8.9Hz, 2H), 7.40 (d, J=8.9Hz, 2H), 3.76 (quintet, J=7.8Hz, 2H), 2.54 (t, J=7.9Hz, 2H),2.43(m,2H),2.11(m,2H).LC-MS:m/z (M+H)=295.0
Step F:N, N'-(6,6'-(Pentamethylene .-1,3-diyl) double (pyridazine-6,3-diyl)) double (2-(pyridine-2-base) Acetamide) (175)
By double for (1R, 3S)-1,3-of stirring (6-chlorine pyridazine-3-base) Pentamethylene. (50mg, 0.16mmol), Cs2CO3 (210mg,0.64mmol)、Pd2(dba)3Double diphenylphosphine-9,9-dimethyl the xanthene of (15mg, 0.016mmol), 4,5- (14mg, 0.024mmol) solution in dioxane was in 100 DEG C of microwave heatings 1 hour.It is cooled to, then filtration over celite Pad, and wash with dioxane.Concentrated filtrate, and by flash chromatography and preparation HPLC purification subsequently, to provide desired Product (15mg, 18%, trans: cis=3:4).
1H NMR(400MHz,CDCl3) cis δ: 8.67 (d, J=4.6Hz, 2H), 8.38 (dd, J=9.1,7.0Hz, 2H), 7.73 (td, J=7.7,1.6Hz, 2H), 7.45 (d, J=9.2Hz, 2H), 7.26-7.29 (m, 4H), 3.96 (s, 4H), 3.60 (m, 2H), 2.61 (dt, J=12.4,6.0Hz, 1H), 2.20-2.42 (m, 3H), 2.10 (m, 2H). trans δ: 8.67 (d, J=4.6Hz, 2H), 8.38 (dd, J=9.1,7.0Hz, 2H), 7.73 (td, J=7.7,1.6Hz, 2H), 7.35 (d, J= 9.2Hz, 2H), 7.26-7.29 (m, 4H), 3.96 (s, 4H), 3.68 (quintet, J=7.6Hz, 2H), 2.37 (t, J= 7.8Hz,2H),2.30(m,2H),2.07(m,2H).LC-MS:m/z (M+H)=494.5
Be thus described several aspects of several embodiment it should be understood that various change, modify and improve right To be easy to perform in those skilled in the art.These change, modify and improve and be intended to as a part disclosed in this, and It is intended to be within the spirit and scope of the present invention.Therefore, aforesaid explanation and graphic being merely possible to are illustrated.

Claims (20)

1. the compound of a formula (I) or its pharmaceutically acceptable salt:
Wherein
X is the C being optionally substituted3-C7Cycloalkylidene；
Each W, Y and Z independently be-S-,-CH=,-CH=CH-,-CH=CR1-、-CR1=CR1-,-O-,-N=or-NH-, Condition be at least one of W, Y and Z for-CH=,
Each W', Y' and Z' independently be-S-,-CH=,-CH=CH-,-CH=CR2-、-CR2=CR2-,-O-,-N=or- NH-, condition is that at least one of W', Y' and Z' is for-CH=；Condition be one of W, Y and Z be-CH=CH-,-CH=CR1- Or-CR1=CR1-, and
When W is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and Z independently be-CH=or-N=；
When Y is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of W and Z independently be-CH=or-N=；
When Z is-CH=CH-,-CH=CR1-or-CR1=CR1In-time, then each of Y and W independently be-CH=or-N=；
Condition be two of W', Y' and Z' for CH=CH-,-CH=CR2-or-CR2=CR2-, and
When W' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, the most each Y' and Z' independently be-CH=or-N=；
When Y' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of W' and Z' independently be-CH=or-N =；
When Z' is-CH=CH-,-CH=CR2-or-CR2=CR2In-time, then each of Y' and W' independently be-CH=or-N =；
Each R1And R2Independently be-NH2、-N(R3)-C(O)-R4、-C(O)-N(R3)-R4、-N(R3)-C(O)-O-R4、-N(R3)- C(O)-N(R3)-R4Or-N (R3)-C(O)-SR4；
Each R3Independently be hydrogen, C1-6Alkyl or aryl；
Each R4Independently be C1-6Alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloalkyl, cycloalkyl-alkyl, heterocycle alkane Base or heterocyclic radical, each of which R occurred by 0 to 3 time5Replace；
Each R5Independently be C1-6Alkyl, C1-6Alkoxyl ,-O-C1-6Alkylidene C1-6Alkoxyl, C1-6Thio alkoxy, C1-6Halogen Substituted alkyl, C3-7Cycloalkyl, C3-7Cycloalkyl-alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloheteroalkylalkyl, heterocyclic radical, Cyano group, halogen, oxo ,-OH ,-OCF3、-OCHF2、-SO2-C1-6Alkyl ,-NO2、-N(R7)-C(O)-C1-6Alkyl ,-C (O) N (R7)2、-N(R7)S(O)1-2-C1-6Alkyl ,-S (O)2N(R7)2、-N(R7)2、-C1-6Alkylidene-N (R7)2, wherein said alkyl, C1-6Alkoxyl ,-O-C1-6Alkylidene C1-6Alkoxyl, C1-6Thio alkoxy, C1-6Haloalkyl, C3-7Cycloalkyl, C3-7Cycloalkyl Alkyl, aryl, heteroaryl, aralkyl, heteroarylalkyl, cycloheteroalkylalkyl, heterocyclic radical ,-SO2-C1-6Alkyl ,-NO2、-N(R7)-C (O)-C1-6Alkyl ,-C (O) N (R7)2、-N(R7)S(O)1-2-C1-6Alkyl ,-S (O)2N(R7)2、-N(R7)2, or-C1-6Alkylidene-N (R7)2The R occurred by 0 to 3 time8Optionally replace；Or two R closed on5Part, atom in connection shape altogether Become cycloalkyl or heterocyclic radical；
Each R6Independently be hydrogen, fluorine, C1-6Alkyl ,-OH ,-NH2、-NH(CH3)、-N(CH3)2, or C1-6Alkoxyl；
Each R7Independently be hydrogen or C1-6Alkyl；
Each R8Independently be halogen, C1-6Alkyl, C1-6Haloalkyl ,-OH ,-N (R7)2, or C1-6Alkoxyl ,-O-C1-6Alkylene Base C1-6Alkoxyl, CN, NO2、-N(R7)-C(O)-C1-6Alkyl ,-C (O) N (R7)2、-N(R7)S(O)1-2C1-6Alkyl or-S (O)2N(R7)2；
M is 0,1 or 2；
N is 0,1 or 2；
O is 1,2 or 3；And
P is 1,2 or 3.
Compound the most according to claim 1, wherein W be-CH=CH-, Y be-N=, Z for-N=, W' for-S-, Y' for- N=and Z' is-N=.
Compound the most according to claim 1, wherein o is 1 and p to be 1.
Compound the most according to claim 1, wherein m is 0 and n to be 0.
Compound the most according to claim 1, wherein n is 1 and m to be 1.
Compound the most according to claim 5, the most each R6It is all hydrogen.
Compound the most according to claim 1, wherein R1And R2Identical.
Compound the most according to claim 1, wherein R1And R2Different.
Compound the most according to claim 1, wherein R1And R2It is respectively-N (R3)-C(O)-R4, the most each R3For hydrogen and Each R4For aralkyl or heteroarylalkyl, each of which R occurred by 0 to 3 time5Replace.
Compound the most according to claim 1, wherein said compound is the compound of formula (II):
11. compounds according to claim 10, wherein said compound is the compound of formula (IIa):
12. compounds according to claim 1, wherein said compound is the compound of formula (IIb):
13. compounds according to claim 1, wherein said compound be the compound of formula (IV) and q be 0,1,2,3 or 4:
14. compounds according to claim 1, wherein said compound is the compound of formula (IVa) and q is 0,1,2,3 Or 4:
15. compounds according to claim 1, wherein said compound is the compound of formula (IVb) and q is 0,1,2,3 Or 4:
16. compounds according to claim 1, wherein said compound is the compound of formula (IVc) and q is 0,1,2,3 Or 4:
The method of the compound of 17. 1 kinds of formulas I
It includes making
Reaction, wherein G is leaving group.
18. 1 kinds of pharmaceutical compositions, its compound comprising formula (I) or its pharmaceutically acceptable salt.
19. 1 kinds of methods treating cancer, described method includes using according to claim 1 to experimenter in need Compound or compositions according to claim 18.
20. methods according to claim 19, wherein said cancer is selected from being characterized as following cancer: i) and reference standard Compare low-level CAM 120/80 expression, ii) high-caliber Vimentin or iii compared with reference standard) low Level or drop low-level carboxylase expression of enzymes.
CN201580020934.4A 2014-03-21 2015-03-20 Compound and its application method Expired - Fee Related CN106231900B (en)

Applications Claiming Priority (3)

Application Number Priority Date Filing Date Title
CNPCT/CN2014/073812 2014-03-21
CN2014073812 2014-03-21
PCT/US2015/021781 WO2015143340A1 (en) 2014-03-21 2015-03-20 Compounds and their methods of use

Publications (2)

Publication Number Publication Date
CN106231900A true CN106231900A (en) 2016-12-14
CN106231900B CN106231900B (en) 2019-05-28

Family

ID=54145391

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201580020934.4A Expired - Fee Related CN106231900B (en) 2014-03-21 2015-03-20 Compound and its application method

Country Status (14)

Country Link
US (1) US10000479B2 (en)
EP (1) EP3119199B1 (en)
JP (2) JP6602364B2 (en)
KR (1) KR20160127149A (en)
CN (1) CN106231900B (en)
AU (1) AU2015231053B2 (en)
BR (1) BR112016021620A2 (en)
CA (1) CA2943339A1 (en)
EA (1) EA201691896A1 (en)
IL (1) IL247921A0 (en)
MX (1) MX2016012244A (en)
PH (1) PH12016501838A1 (en)
WO (1) WO2015143340A1 (en)
ZA (1) ZA201606450B (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110746416A (en) * 2019-09-05 2020-02-04 中国药科大学 Glutaminase GLS1 inhibitor containing triazole structure or pharmaceutically acceptable salt thereof, and preparation method and application thereof

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP2922832B1 (en) 2012-11-21 2019-10-09 Agios Pharmaceuticals, Inc. Glutaminase inhibitors and methods of use
WO2014079011A1 (en) 2012-11-22 2014-05-30 Agios Pharmaceuticals, Inc. Heterocyclic compounds for inhibiting glutaminase and their methods of use
PT3137460T (en) 2014-04-30 2019-12-30 Pfizer Cycloalkyl-linked diheterocycle derivatives

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020115698A1 (en) * 2000-08-10 2002-08-22 Newcomb Robert W. Selective inhibition of glutaminase by bis-thiadiazoles
WO2008048967A1 (en) * 2006-10-16 2008-04-24 University Of Rochester Tripodal cyclohexane derivatives and their use as carbohydrate receptors
WO2013078123A1 (en) * 2011-11-21 2013-05-30 Calithera Biosciences Inc. Heterocyclic inhibitors of glutaminase

Family Cites Families (33)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE2037257A1 (en) 1970-07-28 1972-02-03 Farbwerke Hoechst AG, vorm. Meister Lucius & Brüning, 6000 Frankfurt Poly-(5-amino-1,3,4-thiadiazol-2-yl) derivs prepn - intermediates for drug and polymer prodn
US4070400A (en) 1975-10-16 1978-01-24 Merck & Co., Inc. Diphenyl polyamides having a cyclohexylene moiety
US4376110A (en) 1980-08-04 1983-03-08 Hybritech, Incorporated Immunometric assays using monoclonal antibodies
US4366241A (en) 1980-08-07 1982-12-28 Syva Company Concentrating zone method in heterogeneous immunoassays
US4517288A (en) 1981-01-23 1985-05-14 American Hospital Supply Corp. Solid phase system for ligand assay
CA1291031C (en) 1985-12-23 1991-10-22 Nikolaas C.J. De Jaeger Method for the detection of specific binding agents and their correspondingbindable substances
US4843155A (en) 1987-11-19 1989-06-27 Piotr Chomczynski Product and process for isolating RNA
CA2155759A1 (en) 1993-03-29 1994-10-13 Bernd Janssen 1-amino-3-phenoxy propane derivatives as modulators of multi-drug resistance
US5895748A (en) 1996-11-27 1999-04-20 Johnson; Keith R. Panel of antibodies for detecting cadherins and catenins in tissues and method of using the panel of antibodies
DE69923681T2 (en) 1998-06-18 2006-01-12 Bristol-Myers Squibb Co. CARBON SUBSTITUTED AMINOTHIAZOLE AS INHIBITORS OF CYCLINE-DEPENDENT KINASES
WO2000052204A2 (en) 1999-02-22 2000-09-08 Orntoft Torben F Gene expression in bladder tumors
AU2001249064A1 (en) 2000-02-22 2001-09-03 Mount Sinai School Of Medicine Of New York University N-cadherin modulated migration, invasion, and metastasis
US6974667B2 (en) 2000-06-14 2005-12-13 Gene Logic, Inc. Gene expression profiles in liver cancer
DK1388734T3 (en) 2002-08-01 2004-05-03 Mtm Lab Ag Method for solution-based diagnosis
WO2006040569A1 (en) 2004-10-14 2006-04-20 Astex Therapeutics Limited Thiophene amide compounds for use in the treatment or prophylaxis of cancers
US20080182865A1 (en) 2005-03-11 2008-07-31 Witta Samir E Histone deacetylase inhibitors sensitize cancer cells to epidermal growth factor inhibitors
DE602006016085D1 (en) 2005-03-16 2010-09-23 Genentech Inc BIOLOGICAL MARKERS PREDICTIVE FOR THE APPLICATION OF CANCER TO INHIBITORS OF THE CINEMA OF THE RECEPTOR FOR EPIDERMAL GROWTH FACTOR
CA2702462A1 (en) 2007-11-19 2009-05-28 Genentech, Inc. Compositions and methods for inhibiting tumor progression
JP2011516826A (en) 2008-03-07 2011-05-26 オーエスアイ・フアーマスーテイカルズ・インコーポレーテツド Methods for identification of mesenchymal-like tumor cells or agents that inhibit this formation
US20120190565A1 (en) 2009-02-20 2012-07-26 Pangea Biosciences, Inc. Method Of Diagnosis Or Prognosis Of A Neoplasm Comprising Determining The Level Of Expression Of A Protein In Stromal Cells Adjacent To The Neoplasm
WO2010111504A2 (en) 2009-03-25 2010-09-30 Cornell University Inhibition of glutaminase c
US20120142028A1 (en) 2009-04-17 2012-06-07 OSI Pharmaceuticals, LLC Biological markers predictive of anti-cancer response to epidermal growth factor receptor kinase inhibitors
US20130109643A1 (en) 2010-05-10 2013-05-02 The Johns Hopkins University Metabolic inhibitor against tumors having an idh mutation
JP5944385B2 (en) 2010-06-23 2016-07-05 ユニバーシティ オブ ルーイビル リサーチ ファウンデーション，インコーポレーテッド Method for detecting cancer
US10064885B2 (en) 2010-07-09 2018-09-04 Massachusetts Institute Of Technology Metabolic gene, enzyme, and flux targets for cancer therapy
UY33930A (en) 2011-03-04 2012-10-31 Novartis Ag NEW QUINASE INHIBITORS
CN102827073A (en) * 2011-06-17 2012-12-19 安吉奥斯医药品有限公司 Therapeutically active compositions and application methods thereof
KR20140138956A (en) 2012-03-15 2014-12-04 더 리서치 파운데이션 포 더 스테이트 유니버시티 오브 뉴욕 Combination therapies including inhibitors of the extracellular domain of e-cadherin
EP2922832B1 (en) 2012-11-21 2019-10-09 Agios Pharmaceuticals, Inc. Glutaminase inhibitors and methods of use
WO2014079011A1 (en) 2012-11-22 2014-05-30 Agios Pharmaceuticals, Inc. Heterocyclic compounds for inhibiting glutaminase and their methods of use
US9029531B2 (en) 2012-11-22 2015-05-12 Agios Pharmaceuticals, Inc. Compounds and their methods of use
WO2014138391A1 (en) 2013-03-06 2014-09-12 The Johns Hopkins University Targeting glutamine metabolism in brain tumors
JP6783663B2 (en) * 2014-01-06 2020-11-11 ライゼン・ファーマシューティカルズ・エスアー New glutaminase inhibitor

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20020115698A1 (en) * 2000-08-10 2002-08-22 Newcomb Robert W. Selective inhibition of glutaminase by bis-thiadiazoles
WO2008048967A1 (en) * 2006-10-16 2008-04-24 University Of Rochester Tripodal cyclohexane derivatives and their use as carbohydrate receptors
WO2013078123A1 (en) * 2011-11-21 2013-05-30 Calithera Biosciences Inc. Heterocyclic inhibitors of glutaminase

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN110746416A (en) * 2019-09-05 2020-02-04 中国药科大学 Glutaminase GLS1 inhibitor containing triazole structure or pharmaceutically acceptable salt thereof, and preparation method and application thereof
WO2021042723A1 (en) * 2019-09-05 2021-03-11 中国药科大学 Glutaminase gls1 inhibitor containing triazole structure or pharmaceutically acceptable salt thereof, preparation method therefor, and application thereof

Also Published As

Publication number Publication date
US10000479B2 (en) 2018-06-19
JP6602364B2 (en) 2019-11-06
CN106231900B (en) 2019-05-28
WO2015143340A1 (en) 2015-09-24
CA2943339A1 (en) 2015-09-24
AU2015231053A1 (en) 2016-10-06
BR112016021620A2 (en) 2018-07-10
JP2020019807A (en) 2020-02-06
EP3119199A1 (en) 2017-01-25
KR20160127149A (en) 2016-11-02
ZA201606450B (en) 2019-12-18
US20170137414A1 (en) 2017-05-18
JP2017509703A (en) 2017-04-06
EA201691896A1 (en) 2017-01-30
PH12016501838A1 (en) 2017-01-09
EP3119199B1 (en) 2020-03-18
EP3119199A4 (en) 2017-11-15
MX2016012244A (en) 2017-05-08
IL247921A0 (en) 2016-11-30
AU2015231053B2 (en) 2019-04-11

Similar Documents

Publication Publication Date Title
CN105263915B (en) Glutamine enzyme inhibitor and application method
CN104936954B (en) Compound and its application method
CN102448951B (en) Therapeutic compositions and related methods of use
CN104817498B (en) Therapeutic compound and composition
JP2015078232A (en) Therapeutic compositions and related methods of use
JP5837091B2 (en) Novel N- (4- (azetidine-1-carbonyl) phenyl)-(hetero-) arylsulfonamide derivatives as modulators of pyruvate kinase M2 (PKM2)
CN109415361B (en) Acrylic acid derivative, preparation method thereof and application thereof in medicine
CN109563060A (en) IDO1 inhibitor and its preparation method and application
JP2020019807A (en) Compounds and their methods of use
JP2019508496A (en) TAF1 inhibitors for the treatment of cancer
CN105343073B (en) A kind of compound that can be used as Wnt signal pathway activator and its preparation and application
WO2020179859A1 (en) Pyrrolopyrazole derivative
CN105636943B (en) The chromene derivative of inhibitor as TCR-Nck interaction

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant
TR01 Transfer of patent right
TR01 Transfer of patent right

Effective date of registration: 20210804

Address after: Fa Guoxuleineshi

Patentee after: LES LABORATOIRES SERVIER

Address before: Massachusetts, USA

Patentee before: AGIOS PHARMACEUTICALS Inc.

CF01 Termination of patent right due to non-payment of annual fee
CF01 Termination of patent right due to non-payment of annual fee

Granted publication date: 20190528